U.S. patent number 10,690,674 [Application Number 15/578,874] was granted by the patent office on 2020-06-23 for anti-gitr antibodies for cancer diagnostics.
This patent grant is currently assigned to BRISTOL-MYERS SQUIBB COMPANY. The grantee listed for this patent is BRISTOL-MYERS SQUIBB COMPANY. Invention is credited to Olufemi A. Adelakun, Indrani Chakraborty, Guodong Chen, Michelle Minhua Han, Karla A. Henning, Haichun Huang, Richard Y. Huang, Alan J. Korman, Anne C. Lewin, Huiming Li, Nils Lonberg, Mark J. Selby, Mohan Srinivasan, Changyu Wang, Xi-Tao Wang, Susan Chien-Szu Wong.
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United States Patent |
10,690,674 |
Wang , et al. |
June 23, 2020 |
Anti-GITR antibodies for cancer diagnostics
Abstract
Provided herein are diagnostic antibodies that bind to
glucocorticoid-induced tumor necrosis factor receptor (GITR). Such
antibodies are useful for methods of detecting the expression of
GITR in biological samples, for example, tumor tissue, and
identifying a cancer patient likely to respond to anti-GITR
immunotherapy or predicting whether a cancer patient will respond
to anti-GITR immunotherapy.
Inventors: |
Wang; Xi-Tao (Wellesley,
MA), Adelakun; Olufemi A. (Kendall Park, NJ), Lewin; Anne
C. (Lawrenceville, NJ), Korman; Alan J. (Piedmont,
CA), Selby; Mark J. (San Francisco, CA), Wang;
Changyu (Union City, CA), Huang; Haichun (Fremont,
CA), Henning; Karla A. (Milpitas, CA), Lonberg; Nils
(Woodside, CA), Srinivasan; Mohan (Cupertino, CA), Han;
Michelle Minhua (Piedmont, CA), Chen; Guodong (East
Brunswick, NJ), Huang; Richard Y. (Bridgewater, NJ),
Chakraborty; Indrani (Fremont, CA), Wong; Susan
Chien-Szu (Fremont, CA), Li; Huiming (Lexington,
MA) |
Applicant: |
Name |
City |
State |
Country |
Type |
BRISTOL-MYERS SQUIBB COMPANY |
Princeton |
NJ |
US |
|
|
Assignee: |
BRISTOL-MYERS SQUIBB COMPANY
(Princeton, NJ)
|
Family
ID: |
56194564 |
Appl.
No.: |
15/578,874 |
Filed: |
June 2, 2016 |
PCT
Filed: |
June 02, 2016 |
PCT No.: |
PCT/US2016/035514 |
371(c)(1),(2),(4) Date: |
December 01, 2017 |
PCT
Pub. No.: |
WO2016/196792 |
PCT
Pub. Date: |
December 08, 2016 |
Prior Publication Data
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Document
Identifier |
Publication Date |
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US 20180164321 A1 |
Jun 14, 2018 |
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Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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62170579 |
Jun 3, 2015 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P
43/00 (20180101); A61P 35/00 (20180101); C07K
16/2878 (20130101); C07K 16/2866 (20130101); G01N
33/566 (20130101); G01N 33/57492 (20130101); C07K
2317/24 (20130101); C07K 2317/92 (20130101); G01N
2333/70578 (20130101); G01N 2800/7028 (20130101); C07K
2317/21 (20130101); G01N 2333/4703 (20130101); G01N
2800/52 (20130101); G01N 2333/7151 (20130101); C07K
2317/56 (20130101); G01N 2333/7051 (20130101) |
Current International
Class: |
G01N
33/566 (20060101); G01N 33/574 (20060101); C07K
16/28 (20060101) |
Field of
Search: |
;424/173.1 |
Foreign Patent Documents
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06/078911 |
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Jul 2006 |
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WO |
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2015/031667 |
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Mar 2015 |
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WO |
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2015/187835 |
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Dec 2015 |
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WO |
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Other References
Cohen et al (PLoS ONE; 2010, 5(5): 1-12). cited by examiner .
Baltz, K.M., et al., "Cancer immunoediting by GITR
(glucocorticoid-induced TNF-related protein) ligand in humans: NK
cell/tumor cell interactions," The FASEB Journal, vol.
21(10):2442-2454 (2007). cited by applicant .
Chang, W-C., et al., "Clinical significance of regulatory T cells
and CD8+ effector populations in patients with human endometrial
carcinoma," Cancer, vol. 116(24):5777-5788 (2010). cited by
applicant .
Gasparoto, T. et al., "Patients with oral squamous cell carcinoma
are characterized by increased frequency of suppressive regulatory
T cells in the blood and tumor microenvironment," Cancer
Immunology, Immunotherapy, vol. 59(6):819-828 (2009). cited by
applicant .
International Preliminary Report on Patentability,
PCT/US2016/035514, dated Dec. 5, 2017, 10 pages. cited by applicant
.
International Search Report and Written Opinion, PCT/US2016/035514,
dated Sep. 29, 2016, 15 pages. cited by applicant .
Padovani, C., et al., "Glucocorticoid-induced tumor necrosis factor
receptor expression in patients with cervical human papillomavirus
infection," Revista da Sociedade Brasileira de Medicina Tropical,
vol. 46(3):288-292 (2013). cited by applicant.
|
Primary Examiner: Aeder; Sean E
Attorney, Agent or Firm: Nelson Mullins Riley &
Scarborough LLP Remillard, Esq.; Jane E. Frank; Christopher L.
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a 35 U.S.C. 371 national stage filing of
International Application No. PCT/US2016/035514, filed Jun. 2,
2016, which claims priority to U.S. Provisional Patent Application
Ser. No. 62/170,579, filed Jun. 3, 2015. The contents of the
aforementioned applications are hereby incorporated by reference.
Claims
We claim:
1. A method for determining the level of expression of
glucocorticoid-induced tumor necrosis factor receptor (GITR) on
tumor infiltrating lymphocytes (TILs) and/or tumor cells in a
tissue sample, comprising: (a) contacting a tissue sample from a
patient with an antibody, or antigen-binding portion thereof,
comprising heavy chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 5-7,
respectively, and light chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 8-10,
respectively, wherein the antibody or antigen binding portion
thereof binds to human GITR, (b) detecting the binding of the
antibody to GITR on TILs and/or tumor cells of the tissue sample,
and optionally (c) staining the tissue sample with markers of TILs
and/or tumor cells, and/or with hematoxylin and eosin, to identify
the TILs and/or tumor cells that were identified as GITR positive
in step (b).
2. A method of detecting human glucocorticoid-induced tumor
necrosis factor receptor (GITR) in a biological sample comprising:
(a) contacting the sample with an antibody, or antigen-binding
fragment thereof, comprising heavy chain variable region CDR1,
CDR2, and CDR3 comprising the amino acid sequences set forth in SEQ
ID NOs: 5-7, respectively, and light chain variable region CDR1,
CDR2, and CDR3 comprising the amino acid sequences set forth in SEQ
ID NOs: 8-10, respectively, wherein the antibody or antigen-binding
fragment thereof, binds to human GITR, and (b) detecting the
binding of the antibody to GITR in the sample.
3. The method of claim 1, wherein the sample is a human tissue
sample.
4. The method of claim 3, wherein the human tissue sample is a
tumor tissue sample.
5. The method of claim 2, further comprising a step of determining
the level of GITR protein expression in the sample.
6. The method of claim 4, wherein the tumor tissue sample is a
formalin-fixed paraffin-embedded tumor sample or frozen tumor
tissue.
7. A method of identifying a cancer patient likely to respond to an
anti-GITR immunotherapy comprising: (a) contacting a tumor sample
from the patient with an antibody, or antigen-binding portion
thereof, comprising heavy chain variable region CDR1, CDR2, and
CDR3 comprising the amino acid sequences set forth in SEQ ID NOs:
5-7, respectively, and light chain variable region CDR1, CDR2, and
CDR3 comprising the amino acid sequences set forth in SEQ ID NOs:
8-10, respectively, (b) detecting the binding of the antibody to
GITR in the tumor sample, and (c) determining the level of GITR
protein expression in the sample, wherein a level of GITR protein
above a threshold level indicates that the tumor is a GITR-positive
tumor and that the patient is likely to or predicted to respond to
anti-GITR immunotherapy.
8. The method of claim 7, further comprising a step of
administering to the patient an anti-GITR immunotherapy if the
tumor is determined to be GITR-positive.
9. A method of treating a cancer patient comprising: (a)
determining whether the patient has a GITR-positive tumor according
to the method of claim 2; and (b) administering to the patient an
agent that modulates GITR signaling if the patient is determined to
have a GITR-positive tumor.
10. The method of claim 8, wherein the agent is an agonistic
anti-GITR antibody.
11. A method of monitoring a glucocorticoid-induced tumor necrosis
factor receptor (GITR)-positive tumor in a cancer patient
comprising: (a) detecting GITR protein expression in the
GITR-positive tumor at a first time point by using an antibody
comprising heavy chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 5-7,
respectively, and light chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 8-10,
respectively, (b) determining the level of GITR protein expression
at the first time point, (c) detecting GITR protein expression in
the tumor at a second time point using the same antibody used in
step (a), (d) determining the level of GITR protein expression in
the tumor from the second time point, and (e) comparing the levels
of GITR protein expression determined at the first and second time
points, wherein a higher level at the first time point relative to
the second time point is indicative of tumor regression, a lower
score at the first time point relative to the second time point is
indicative of tumor progression, and a relatively unchanged level
at the first time point relative to the second time point is
indicative of a stable tumor.
12. The method of claim 1, wherein the antibody comprises heavy and
light chain variable regions comprising the amino acid sequences
set forth in SEQ ID NOs: 11 and 12, respectively.
13. The method of claim 1, wherein the antibody is an IgG1, IgG2,
IgG3, or IgG4 antibody.
14. The method of claim 1, wherein the antibody is a humanized,
human, or chimeric antibody.
15. The method of claim 14, wherein the antibody is a chimeric
antibody which has human variable region sequences and non-human
constant region sequences.
16. The method of claim 15, wherein the non-human constant region
comprises a mouse Fc region.
17. The method of claim 16, wherein the mouse Fc region is a mouse
IgG2a Fc region.
18. The method of claim 15, wherein the chimeric antibody comprises
heavy and light chain sequences set forth in SEQ ID NOs: 15 and 16,
respectively.
19. The method of claim 2, wherein the antibody, or antigen-binding
portion thereof, comprises a detectable moiety.
20. A diagnostic kit comprising an antibody, or antigen-binding
portion thereof, comprising heavy chain variable region CDR1, CDR2,
and CDR3 comprising the amino acid sequences set forth in SEQ ID
NOs: 5-7, respectively, and light chain variable region CDR1, CDR2,
and CDR3 comprising the amino acid sequences set forth in SEQ ID
NOs: 8-10, respectively, and instructions for use.
21. The method of claim 2, wherein the antibody comprises heavy and
light chain variable regions comprising the amino acid sequences
set forth in SEQ ID NOs: 11 and 12, respectively.
22. The method of claim 2, wherein the antibody comprises a mouse
IgG2a Fc region.
23. The method of claim 2, wherein the antibody comprises heavy and
light chain sequences set forth in SEQ ID NOs: 15 and 16,
respectively.
24. The kit of claim 20, wherein the antibody comprises heavy and
light chain variable regions comprising the amino acid sequences
set forth in SEQ ID NOs: 11 and 12, respectively.
25. The kit of claim 20, wherein the antibody comprises a mouse
IgG2a Fc region.
26. The kit of claim 20, wherein the antibody comprises heavy and
light chain sequences set forth in SEQ ID NOs: 15 and 16,
respectively.
27. The kit of claim 20, wherein the antibody, or antigen-binding
portion thereof, comprises a detectable moiety.
Description
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been
submitted electronically in ASCII format and is hereby incorporated
by reference in its entirety. Said ASCII copy, created on Dec. 1,
2017, is named MXI_544US_Sequence_Listing.txt and is 485,984 bytes
in size.
BACKGROUND
Glucocorticoid-inducible TNF receptor (GITR) is a T cell
costimulatory type I cell surface transmembrane glycoprotein
expressed by activated effector T (Teff) cells and at low levels on
B cells, eosinophils, basophils, and macrophages (Nocentini et al.,
Adv Exp Med Biol 2009; 647:156-73). GITR is constitutively
expressed on regulatory T (Treg) cells. Signaling through GITR
potentiates T cell receptor activation of T cells and renders Teff
cells resistant to Treg-mediated suppression (Stephens et al., J
Immunol 2004; 173:5008-20). Determining GITR positivity in tumor
tissue may provide an opportunity to target patients that are most
likely to benefit from (i.e., be responsive to) anti-GITR
immunotherapy, as well as monitoring the status of GITR-positive
tumors and efficacy of anti-GITR immunotherapy.
SUMMARY
Provided herein are diagnostic anti-GITR antibodies and methods for
detecting GITR protein expression in biological samples, such as
tumor tissue samples. Such methods may be useful for, e.g.,
identifying cancer patients who are likely to respond to anti-GITR
immunotherapy and monitoring the status of GITR-positive tumors and
efficacy of anti-GITR immunotherapy.
Accordingly, in one aspect, provided herein are methods for
detecting human GITR in a biological sample, for example, a human
tissue sample (e.g., tumor tissue sample), comprising contacting
the sample with an antibody, or antigen-binding fragment thereof,
comprising heavy chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 5-7,
respectively, and light chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 8-10,
respectively, wherein the antibody or antigen-binding fragment
thereof specifically binds to human GITR, and detecting the binding
of the antibody to GITR in the sample.
Also provided herein are methods of determining whether a tumor of
a cancer patient is GITR-positive comprising:
(a) contacting a tumor sample from the patient with an antibody, or
antigen-binding portion thereof, comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human
GITR,
(b) detecting the binding of the antibody to GITR in the sample,
and
(c) determining the level of GITR protein expression in the sample,
wherein a level of GITR protein above a threshold level indicates
that the tumor is GITR-positive.
Also provided herein are methods of identifying a cancer patient
likely to respond to an anti-GITR immunotherapy comprising:
(a) contacting a tumor sample from the patient with an antibody, or
antigen-binding portion thereof, comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human
GITR,
(b) detecting the binding of the antibody to GITR in the tumor
sample,
(c) determining the level of GITR protein expression in the sample,
wherein a level of GITR protein above a threshold level indicates
that the tumor is GITR-positive tumor and that the patient is
likely to or predicted to respond to anti-GITR immunotherapy.
Also provide herein are methods of treating a cancer patient
comprising:
(a) determining whether a tumor of a cancer patient is
GITR-positive by (i) contacting a tumor sample from the patient
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR, (ii) detecting the binding of the
antibody to GITR in the tumor sample, and (iii) determining the
level of GITR protein expression in the sample, wherein a level of
GITR protein above a threshold level indicates that the tumor is
GITR-positive, and
(b) if the tumor is determined to be GITR-positive, administering
to the patient an agent that modulates GITR signaling. In certain
embodiments, the agent that modulates GITR signaling is an
agonistic (i.e., agonist) anti-GITR antibody.
Also provided herein are methods of monitoring a GITR-positive
tumor in a cancer patient comprising:
(a) detecting GITR protein expression in the GITR-positive tumor at
a first time point by using an antibody comprising heavy chain
variable region CDR1, CDR2, and CDR3 comprising the amino acid
sequences set forth in SEQ ID NOs: 5-7, respectively, and light
chain variable region CDR1, CDR2, and CDR3 comprising the amino
acid sequences set forth in SEQ ID NOs: 8-10, respectively, wherein
the antibody or antigen-binding fragment thereof specifically binds
to human GITR,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in the tumor at a second time
point using the same antibody used in step (a),
(d) determining the level of GITR protein expression in the tumor
from the second time point;
(e) comparing the levels of GITR protein expression determined at
the first and second time points, wherein a higher level at the
first time point relative to the second time point is indicative of
tumor regression, a lower score at the first time point relative to
the second time point is indicative of tumor progression, and a
relatively unchanged level at the first time point relative to the
second time point is indicative of a stable tumor.
Also provided herein are methods of monitoring the efficacy of
anti-GITR immunotherapy in a patient having a GITR-positive tumor
comprising:
(a) detecting GITR protein expression in the GITR-positive tumor at
a first time point before or after initiating anti-GITR
immunotherapy by using an antibody comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human
GITR,
(b) determining the level of GITR protein expression in the tumor
from the first time point,
(c) detecting GITR protein expression in the GITR-positive tumor at
a second time point after initiating anti-GITR immunotherapy using
the same antibody from step (a),
(d) determining the level of GITR protein expression in the tumor
from the second time point;
(e) comparing the levels of GITR protein expression determined at
the first and second time points, wherein a higher level at the
first time point relative to the second time point is indicative of
effective anti-GITR immunotherapy, a lower score at the first time
point relative to the second time point is indicative of
ineffective anti-GITR immunotherapy, and an unchanged score at the
first time point relative to the second time point is indicative of
anti-GITR immunotherapy being stabilizing. In certain embodiments,
the anti-GITR immunotherapy comprises administering to the patient
a therapeutically effective amount of an anti-GITR antibody, or
antigen binding portion thereof.
In certain embodiments, the tumor sample used in the methods
described herein is a formalin-fixed paraffin-embedded tumor
sample. In certain embodiments, the tumor sample is a frozen tumor
tissue sample. In certain embodiments, the methods described herein
are performed by immunohistochemistry. In certain embodiments, the
level of GITR expression is determined using a scoring system,
e.g., the H score system, the Allred score system, the AQUA system,
or an automated scoring system.
In certain embodiments, the antibody used in the methods described
herein comprises heavy and light chain variable regions comprising
the amino acid sequences set forth in SEQ ID NOs: 11 and 12,
respectively. In certain embodiments, the antibody used in the
methods described herein comprises heavy and light chains
comprising the amino acid sequences set forth in SEQ ID NOs: 13 and
14, respectively. In certain embodiments, the heavy chain lacks the
C-terminal lysine. In certain embodiments, the antibody is an IgG1,
IgG2, IgG3, or IgG4 antibody. In certain embodiments, the antibody
binds to soluble human GITR with a K.sub.D of 10 nM or less, as
assessed by Biacore.
In certain embodiments, the antibody is a humanized, human, or
chimeric antibody. In certain embodiments, the antibody is a
chimeric antibody comprising human variable region sequences and
non-human constant region sequences, for example, a mouse heavy and
light chain constant region. In certain embodiments, the mouse Fc
region is a mouse IgG2a Fc region. In certain embodiments, the
chimeric antibody comprises heavy and light chain sequences set
forth in SEQ ID NOs: 15 and 16, respectively. In certain
embodiments, the heavy chain lacks the C-terminal lysine.
In certain embodiments, the antibody used in the methods described
herein comprises a detectable moiety, such as a radiolabel,
fluorescent label, an enzymatic label, biotin, chromophore, or an
ECL label.
Also provided herein are diagnostic kits comprising an antibody, or
antigen-binding portion thereof, comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, wherein the antibody or antigen-binding
fragment thereof specifically binds to human GITR, and instructions
for use. In certain embodiments, the antibody, or antigen-binding
portion thereof, comprises the heavy and light chain variable
region sequences set forth in SEQ ID NOs: 11 and 12, respectively.
In certain embodiments, the antibody comprises the heavy and light
chain sequences set forth in SEQ ID NOs: 13 and 14, respectively.
In certain embodiments, the antibody comprises the heavy and light
chain sequences set forth in SEQ ID NOs: 15 and 16, respectively.
In certain embodiments, the heavy chain lacks the C-terminal
lysine.
BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 shows the nucleotide sequence (SEQ ID NO: 17) and amino acid
sequence (SEQ ID NO: 11) of the heavy chain variable region of the
6G10 human monoclonal antibody. The CDR1 (SEQ ID NO: 5), CDR2 (SEQ
ID NO: 6) and CDR3 (SEQ ID NO: 7) regions are delineated and the V,
D and J germline derivations are indicated.
FIG. 2 shows the nucleotide sequence (SEQ ID NO: 18) and amino acid
sequence (SEQ ID NO: 12) of the kappa light chain variable region
of the 6G10 human monoclonal antibody. The CDR1 (SEQ ID NO: 8),
CDR2 (SEQ ID NO: 9) and CDR3 (SEQ ID NO: 10) regions are delineated
and the V and J germline derivations are indicated.
FIG. 3 shows an alignment of the amino acid sequence of the heavy
chain variable regions of 6G10 (SEQ ID NO: 11) with the human
germline V.sub.H 3-33, 3-10, and JH6 amino acid sequences.
FIG. 4 shows an alignment of the amino acid sequence of the light
chain variable region (VK1) of 6G10 (SEQ ID NO: 12) with the human
germline V.sub.k L18 and JK2 amino acid sequences.
FIG. 5 shows the nucleotide sequence (SEQ ID NO: 21) and amino acid
sequence (SEQ ID NO: 15) of the heavy chain of the chimeric 6G10
antibody.
FIG. 6 shows the nucleotide sequence (SEQ ID NO: 22) and amino acid
sequence (SEQ ID NO: 16) of the light chain of the chimeric 6G10
antibody.
FIG. 7 shows staining of CD3, FoxP3, and GITR on adjacent
formalin-fixed paraffin embedded (FFPE) sections from cervical
squamous cell carcinoma samples by immunohistochemistry using the
chimeric 6G10 antibody.
DETAILED DESCRIPTION
Disclosed herein are various methods for detecting GITR expression
in biological samples, e.g., tumor specimens, using diagnostic
anti-GITR antibodies. Such antibodies may be useful for, e.g.,
monitoring GITR protein levels in a subject; identifying the
specific cells that are GITR positive in a specific tissue, such as
a tumor tissue (e.g., identifying the presence of GITR in Treg
cells and/or tumor cells); the early detection of cancers;
monitoring and identification of existing cancers; and assessing
the therapeutic efficacy of anti-GITR immunotherapy.
Definitions
In order that the present description may be more readily
understood, certain terms are first defined. Additional definitions
are set forth throughout the detailed description.
The term "glucocorticoid-inducible TNF receptor" or "GITR" as used
herein refers to a receptor that is a member of the TNF-receptor
superfamily, which binds to GITR ligand (GITR-L). GITR is also
referred to as tumor necrosis factor receptor superfamily, member
18 (TNFRSF18), AITR and CD357. The term "GITR" includes any
variants or isoforms of GITR which are naturally expressed by
cells. Accordingly, antibodies described herein may cross-react
with GITR from species other than human (e.g., cynomolgus GITR).
Alternatively, the antibodies may be specific for human GITR and
may not exhibit any cross-reactivity with other species. GITR or
any variants and isoforms thereof, may either be isolated from
cells or tissues which naturally express them or be recombinantly
produced using well-known techniques in the art and/or those
described herein.
Three isoforms of human GITR have been identified, all of which
share the same extracellular domain, except for its C-terminal
portion. Variant 1 (Accession No. NP_004186; SEQ ID NO: 1) consists
of 241 amino acids and represents the longest transcript. It
contains an extra coding segment that leads to a frame shift,
compared to variant 2. The resulting protein (isoform 1) contains a
distinct and shorter C-terminus, as compared to isoform 2. Variant
2 (Accession No. NP_683699; SEQ ID NO: 2) encodes the longest
protein (isoform 2), consisting of 255 amino acids, and is soluble.
Variant 3 (Accession No. NP_683700; SEQ ID NO: 3) contains an extra
coding segment that leads to a frame shift, compared to variant 2.
The resulting protein (isoform 3) contains a distinct and shorter
C-terminus, as compared to isoform 2, and consists of 234 amino
acids. The amino acid sequences of human GITR isoforms 1, 2, and 3
are set forth in SEQ ID NOs: 1, 2, and 3, respectively, and the
extracellular domain of mature GITR has the amino acid sequence set
forth in SEQ ID NO: 4.
The term "antibody" as used to herein includes whole antibodies and
any antigen binding fragments (i.e., "antigen-binding portions") or
single chains thereof. An "antibody" refers, in one embodiment, to
a glycoprotein comprising at least two heavy (H) chains and two
light (L) chains inter-connected by disulfide bonds, or an antigen
binding portion thereof. Each heavy chain is comprised of a heavy
chain variable region (abbreviated herein as V.sub.H) and a heavy
chain constant region. In certain naturally occurring antibodies,
the heavy chain constant region is comprised of three domains, CH1,
CH2 and CH3. In certain naturally occurring antibodies, each light
chain is comprised of a light chain variable region (abbreviated
herein as V.sub.L) and a light chain constant region. The light
chain constant region is comprised of one domain, CL. The V.sub.H
and V.sub.L regions can be further subdivided into regions of
hypervariability, termed complementarity determining regions (CDR),
interspersed with regions that are more conserved, termed framework
regions (FR). Each V.sub.H and V.sub.L is composed of three CDRs
and four FRs, arranged from amino-terminus to carboxy-terminus in
the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The
variable regions of the heavy and light chains contain a binding
domain that interacts with an antigen. The constant regions of the
antibodies may mediate the binding of the immunoglobulin to host
tissues or factors, including various cells of the immune system
(e.g., effector cells) and the first component (Clq) of the
classical complement system.
Antibodies typically bind specifically to their cognate antigen
with high affinity, reflected by a dissociation constant (K.sub.D)
of 10.sup.-5 to 10.sup.-11 M or less. Any K.sub.D greater than
about 10.sup.-4 M is generally considered to indicate nonspecific
binding. As used herein, an antibody that "binds specifically" to
an antigen refers to an antibody that binds to the antigen and
substantially identical antigens with high affinity, which means
having a K.sub.D of 10.sup.-7 M or less, preferably 10.sup.-8 M or
less, even more preferably 5.times.10.sup.-9 M or less, and most
preferably between 10.sup.-8 M and 10.sup.-10 M or less, but does
not bind with high affinity to unrelated antigens.
An immunoglobulin may be from any of the commonly known isotypes,
including but not limited to IgA, secretory IgA, IgG and IgM. The
IgG isotype is divided in subclasses in certain species: IgG1,
IgG2, IgG3 and IgG4 in humans, and IgG1, IgG2a, IgG2b and IgG3 in
mice. In certain embodiments, the anti-GITR antibodies described
herein are of the IgG1 or IgG2 subtype. Immunoglobulins, e.g.,
IgG1, exist in several allotypes, which differ from each other in
at most a few amino acids. "Antibody" includes, by way of example,
both naturally occurring and non-naturally occurring antibodies;
monoclonal and polyclonal antibodies; chimeric and humanized
antibodies; human and nonhuman antibodies; wholly synthetic
antibodies; and single chain antibodies.
The term "antigen-binding portion" of an antibody, as used herein,
refers to one or more fragments of an antibody that retain the
ability to specifically bind to an antigen (e.g., human GITR). Such
"fragments" are, for example between about 8 and about 1500 amino
acids in length, suitably between about 8 and about 745 amino acids
in length, suitably about 8 to about 300, for example about 8 to
about 200 amino acids, or about 10 to about 50 or 100 amino acids
in length. It has been shown that the antigen-binding function of
an antibody can be performed by fragments of a full-length
antibody. Examples of binding fragments encompassed within the term
"antigen-binding portion" of an antibody, e.g., an anti-GITR
antibody described herein, include (i) a Fab fragment, a monovalent
fragment consisting of the V.sub.L, V.sub.H, CL and CH1 domains;
(ii) a F(ab').sub.2 fragment, a bivalent fragment comprising two
Fab fragments linked by a disulfide bridge at the hinge region;
(iii) a Fd fragment consisting of the V.sub.H and CH1 domains; (iv)
a Fv fragment consisting of the V.sub.L and V.sub.H domains of a
single arm of an antibody, (v) a dAb fragment (Ward et al., (1989)
Nature 341:544-546), which consists of a V.sub.H domain; and (vi)
an isolated complementarity determining region (CDR) or (vii) a
combination of two or more isolated CDRs which may optionally be
joined by a synthetic linker. Furthermore, although the two domains
of the Fv fragment, V.sub.L and V.sub.H, are coded for by separate
genes, they can be joined, using recombinant methods, by a
synthetic linker that enables them to be made as a single protein
chain in which the V.sub.L and V.sub.H regions pair to form
monovalent molecules (known as single chain Fv (scFv); see e.g.,
Bird et al. (1988) Science 242:423-426; and Huston et al. (1988)
Proc. Natl. Acad. Sci. USA 85:5879-5883). Such single chain
antibodies are also intended to be encompassed within the term
"antigen-binding portion" of an antibody. These antibody fragments
are obtained using conventional techniques known to those with
skill in the art, and the fragments are screened for utility in the
same manner as are intact antibodies. Antigen-binding portions can
be produced by recombinant DNA techniques, or by enzymatic or
chemical cleavage of intact immunoglobulins.
The term "monoclonal antibody," as used herein, refers to an
antibody that displays a single binding specificity and affinity
for a particular epitope or a composition of antibodies in which
all antibodies display a single binding specificity and affinity
for a particular epitope. Accordingly, the term "human monoclonal
antibody" refers to an antibody or antibody composition that
display(s) a single binding specificity and which has variable and
optional constant regions derived from human germline
immunoglobulin sequences. In one embodiment, human monoclonal
antibodies are produced by a hybridoma which includes a B cell
obtained from a transgenic non-human animal, e.g., a transgenic
mouse, having a genome comprising a human heavy chain transgene and
a light chain transgene fused to an immortalized cell.
The term "recombinant human antibody," as used herein, includes all
human antibodies that are prepared, expressed, created or isolated
by recombinant means, such as (a) antibodies isolated from an
animal (e.g., a mouse) that is transgenic or transchromosomal for
human immunoglobulin genes or a hybridoma prepared therefrom, (b)
antibodies isolated from a host cell transformed to express the
antibody, e.g., from a transfectoma, (c) antibodies isolated from a
recombinant, combinatorial human antibody library, and (d)
antibodies prepared, expressed, created or isolated by any other
means that involve splicing of human immunoglobulin gene sequences
to other DNA sequences. Such recombinant human antibodies comprise
variable and constant regions that utilize particular human
germline immunoglobulin sequences are encoded by the germline
genes, but include subsequent rearrangements and mutations which
occur, for example, during antibody maturation. As known in the art
(see, e.g., Lonberg, Nature Biotech 2005; 23:1117-25), the variable
region contains the antigen binding domain, which is encoded by
various genes that rearrange to form an antibody specific for a
foreign antigen. In addition to rearrangement, the variable region
can be further modified by multiple single amino acid changes
(referred to as somatic mutation or hypermutation) to increase the
affinity of the antibody to the foreign antigen. The constant
region will change in further response to an antigen (i.e., isotype
switch). Therefore, the rearranged and somatically mutated nucleic
acid molecules that encode the light chain and heavy chain
immunoglobulin polypeptides in response to an antigen may not have
sequence identity with the original nucleic acid molecules, but
instead will be substantially identical or similar (i.e., have at
least 80% identity).
A "human" antibody (HuMAb) refers to an antibody having variable
regions in which both the framework and CDR regions are derived
from human germline immunoglobulin sequences. Furthermore, if the
antibody contains a constant region, the constant region also is
derived from human germline immunoglobulin sequences. The
antibodies described herein may include amino acid residues not
encoded by human germline immunoglobulin sequences (e.g., mutations
introduced by random or site-specific mutagenesis in vitro or by
somatic mutation in vivo). However, the term "human antibody", as
used herein, is not intended to include antibodies in which CDR
sequences derived from the germline of another mammalian species,
such as a mouse, have been grafted onto human framework sequences.
The terms "human" antibodies and "fully human" antibodies and are
used synonymously.
A "humanized" antibody refers to an antibody in which some, most or
all of the amino acids outside the CDR domains of a non-human
antibody are replaced with corresponding amino acids derived from
human immunoglobulins. In one embodiment of a humanized form of an
antibody, some, most or all of the amino acids outside the CDR
domains have been replaced with amino acids from human
immunoglobulins, whereas some, most or all amino acids within one
or more CDR regions are unchanged. Small additions, deletions,
insertions, substitutions or modifications of amino acids are
permissible as long as they do not abrogate the ability of the
antibody to bind to a particular antigen. A "humanized" antibody
retains an antigenic specificity similar to that of the original
antibody.
A "chimeric antibody" refers to an antibody in which the variable
regions are derived from one species (e.g., human variable regions)
and the constant regions are derived from another species (e.g.,
mouse constant regions), such as an antibody in which the variable
regions are derived from a mouse antibody and the constant regions
are derived from a human antibody. Constant region sequences for
various species are known in the art and could be readily obtained
by the skilled artisan to generate chimeric antibodies. An
exemplary chimeric anti-GITR antibody comprises human variable
region sequences and mouse constant chain sequences. For example,
in one embodiment, a chimeric anti-GITR antibody comprises the
heavy and light chain sequences set forth in SEQ ID NOs: 15 and 16,
respectively. In certain embodiments, the heavy chain of the
chimeric antibody lacks the C-terminal lysine.
An "isolated antibody," as used herein, is intended to refer to an
antibody which is substantially free of other antibodies having
different antigenic specificities (e.g., an isolated antibody that
specifically binds to GITR is substantially free of antibodies that
specifically bind antigens other than GITR). An isolated antibody
that specifically binds to an epitope of GITR may, however, have
cross-reactivity to other GITR proteins from different species.
As used herein, the terms "specific binding," "selective binding,"
"selectively binds," and "specifically binds," refer to antibody
binding to an epitope on a predetermined antigen. Typically, the
antibody (i) binds with an equilibrium dissociation constant
(K.sub.D) of approximately less than 10.sup.-7 M, such as
approximately less than 10.sup.-8 M, 10.sup.-9 M or 10.sup.-10 M or
even lower when determined by, e.g., surface plasmon resonance
(SPR) technology in a BIACORE 2000 instrument using the
predetermined antigen, e.g., recombinant human GITR, as the analyte
and the antibody as the ligand.
The term "k.sub.assoc" or "k.sub.a", as used herein, is intended to
refer to the association rate of a particular antibody-antigen
interaction, whereas the term "k.sub.dis" or "k.sub.d," as used
herein, is intended to refer to the dissociation rate of a
particular antibody-antigen interaction. The term "K.sub.D", as
used herein, is intended to refer to the dissociation constant,
which is obtained from the ratio of k.sub.d to k.sub.a (i.e.,
k.sub.d/k.sub.a) and is expressed as a molar concentration (M).
K.sub.D values for antibodies can be determined using methods well
established in the art. A preferred method for determining the
K.sub.D of an antibody is by using surface plasmon resonance,
preferably using a biosensor system such as a Biacore.RTM.
system.
A "polypeptide" refers to a chain comprising at least two
consecutively linked amino acid residues, with no upper limit on
the length of the chain. One or more amino acid residues in the
protein may contain a modification such as, but not limited to,
glycosylation, phosphorylation or disulfide bond formation. A
"protein" may comprise one or more polypeptides.
"Immunotherapy" refers to the treatment of a subject afflicted
with, or at risk of contracting or suffering a recurrence of, a
disease by a method comprising inducing, enhancing, suppressing or
otherwise modifying an immune response. In this context, "anti-GITR
immunotherapy" refers to the treatment of a subject afflicted with,
or at risk of contracting or suffering a recurrence of, a disease
(e.g., cancer) by a method comprising inducing, enhancing,
suppressing or otherwise modifying an immune response by
administering an agent (e.g., an agonistic anti-GITR antibody) that
modulates GITR signaling and results in, e.g., the induction and/or
enhancement of T cell activation (e.g., increase in IL-2 and/or
IFN-.gamma. production by T cells and/or increase in proliferation
of T cells), depletion of regulatory T cells, etc.
As used herein, the term "linked" refers to the association of two
or more molecules. The linkage can be covalent or non-covalent. The
linkage also can be genetic (i.e., recombinantly fused). Such
linkages can be achieved using a wide variety of art recognized
techniques, such as chemical conjugation and recombinant protein
production.
As used herein, "administering" refers to the physical introduction
of a composition comprising a therapeutic agent to a subject, using
any of the various methods and delivery systems known to those
skilled in the art. Preferred routes of administration for
antibodies described herein include intravenous, intraperitoneal,
intramuscular, subcutaneous, spinal or other parenteral routes of
administration, for example by injection or infusion. The phrase
"parenteral administration" as used herein means modes of
administration other than enteral and topical administration,
usually by injection, and includes, without limitation,
intravenous, intraperitoneal, intramuscular, intraarterial,
intrathecal, intralymphatic, intralesional, intracapsular,
intraorbital, intracardiac, intradermal, transtracheal,
subcutaneous, subcuticular, intraarticular, subcapsular,
subarachnoid, intraspinal, epidural and intrasternal injection and
infusion, as well as in vivo electroporation. Alternatively, an
antibody described herein can be administered via a non-parenteral
route, such as a topical, epidermal or mucosal route of
administration, for example, intranasally, orally, vaginally,
rectally, sublingually or topically. Administering can also be
performed, for example, once, a plurality of times, and/or over one
or more extended periods.
As used herein, "cancer" refers to a broad group of diseases
characterized by the uncontrolled growth of abnormal cells in the
body. Unregulated cell division may result in the formation of
malignant tumors or cells that invade neighboring tissues and may
metastasize to distant parts of the body through the lymphatic
system or bloodstream.
As used herein, a "tumor" refers to an abnormal mass of tissue
resulting from excessive cell division that is uncontrolled and
progressive. Tumors can be benign or malignant.
The term "patient" includes human and other mammalian subjects that
receive either prophylactic or therapeutic treatment.
As used herein, the term "subject" includes any human or non-human
animal. For example, the methods and compositions described herein
can be used to treat a subject having cancer. The term "non-human
animal" includes all vertebrates, e.g., mammals and non-mammals,
such as non-human primates, sheep, dog, cow, chickens, amphibians,
reptiles, etc. Various aspects described herein are described in
further detail in the following subsections.
As used herein, the singular forms "a," "an," and "the" include
plural referents unless the context clearly dictates otherwise. The
use of "or" or "and" means "and/or" unless stated otherwise.
Furthermore, use of the term "including" as well as other forms,
such as "include," "includes," and "included," is not limiting.
The term "about" as used herein when referring to a measurable
value such as an amount, temporal duration, and the like,
encompasses variations of up to .+-.10% from the specified value.
Unless otherwise indicated, all numbers expressing quantities of
ingredients, properties such as molecular weight, reaction
conditions, scores in an art-recognized scoring system, etc., used
herein are understood as being modified by the term "about."
The term "sample of a subject" refers to cells (e.g., tumor cells,
T cells, B cells or other), tissues (e.g., tumor), fluids and/or
combination thereof isolated from a subject.
The term "control sample" or "reference sample" as used herein
refers to any clinically relevant control or reference sample,
including, e.g., a sample from a healthy subject or a sample made
at an earlier time from the subject being assessed. For example, a
control sample or reference sample can be a sample taken from a
subject prior to onset of cancer, an earlier stage of disease, or
before administration of treatment.
As used herein, "threshold level" or "cut-off level" in the context
of expression levels refers to a level of GITR protein expression
above which a patient sample (e.g., a patient tumor sample) is
deemed "GITR-positive" and below which the sample is deemed
"GITR-negative." The threshold level may be based on, e.g., one or
more compilations of data from "normal" patient samples (e.g., a
patient population who do not have GITR-positive tumors) or
matching normal tissue from the patient being assessed. The
threshold level may be placed in the framework of an art-recognized
scoring system (e.g., the H score system). For example, in one
embodiment, the tumor of a patient having an H score of 1 in
matching normal tissue would be considered GITR-positive if the H
score of the tumor is greater than 1. A threshold detection level
may be the lowest detection level using a given detection method.
For example, a threshold level may be the level below which GITR
cannot be detected using immunohistochemistry, e.g., using an
immunuohistochemistry method described herein.
Diagnostic Anti-GITR Antibodies
Provided herein are diagnostic anti-GITR antibodies that are useful
for detecting GITR protein expression in human tissue, e.g., tumor
tissue. Such antibodies are useful, e.g., for identifying/selecting
subjects who are likely to benefit from (i.e., be responsive to)
treatment with anti-GITR immunotherapy, monitoring the progression
of a GITR-positive tumor, and/or monitoring the efficacy of
anti-GITR immunotherapy.
Accordingly, in certain embodiments, a diagnostic anti-GITR
antibody useful for detecting GITR protein expression in tissue
samples, e.g., human tumor tissue samples, comprises the heavy and
light chain variable region CDR sequences, heavy and light chain
variable region sequences, or full length heavy and light chain
sequences of antibody 6G10 (a fully human anti-GITR antibody). A
summary of the relevant sequences for 6G10 is provided in Table
2.
In some embodiments, the anti-GITR antibody has the heavy chain
variable region CDR1, CDR2, and CDR3 sequences set forth in SEQ ID
NOs: 5-7, and the light chain variable region CDR1, CDR2, and CDR3
sequences set forth in SEQ ID NOs: 8-10, and specifically binds to
human GITR.
In some embodiments, the anti-GITR antibody has the heavy chain
variable region sequence set forth in SEQ ID NO: 11, and the light
chain variable region sequence set forth in SEQ ID NO: 12.
In some embodiments, the anti-GITR antibody has heavy chain and
light chain sequences set forth in SEQ ID NOs: 13 and 14,
respectively. In certain embodiments, the heavy chain lacks the
C-terminal lysine.
In some embodiments, the anti-GITR antibody is an IgG1, IgG2, IgG3,
or IgG4 antibody, or a variant thereof. In other embodiments, the
antibody is an IgA, IgD, IgE, or IgM antibody.
In some embodiments, the anti-GITR antibody is a human antibody, a
humanized antibody, or a chimeric antibody.
In some embodiments, the anti-GITR antibody binds to soluble GITR
with a K.sub.D of 10 nM or less as determined by Biacore.
In certain embodiments, the anti-GITR antibody is a chimeric
antibody. In one embodiment, the anti-GITR antibody has the heavy
and light chain variable region sequences of the 6G10 antibody (SEQ
ID NOs: 11 and 12, respectively), and the constant regions of a
non-human species, e.g., a mouse IgG2a constant region (SEQ ID NO:
25). Such chimeric 6G10 antibodies are useful for, e.g., reducing
background staining in immunohistochemistry when detecting GITR
protein expression in human tissue. Accordingly, in one embodiment,
the diagnostic anti-GITR antibody is a chimeric antibody comprising
the heavy and light chain sequences set forth in SEQ ID NOs: 15 and
16, respectively. In certain embodiments, the heavy chain of the
chimeric antibody lacks the C-terminal lysine.
In certain embodiments, the diagnostic anti-GITR antibodies
described herein comprise a detectable moiety to facilitate
detection of GITR expression in, for example, tumor samples by,
e.g., immunohistochemistry, as described infra. Exemplary detection
moieties include luminescent labels, fluorescent labels,
radiolabels (e.g., .sup.99Tc, .sup.45Ti, .sup.112In, .sup.111In,
.sup.3H, .sup.121I, .sup.125I, .sup.131I, .sup.14C, .sup.18F,
.sup.36Cl, .sup.55Co, .sup.58Co, .sup.51Cr, .sup.67Cu, .sup.64Cu,
.sup.66Ga, .sup.68Ga, .sup.76Br, .sup.89Zr, .sup.35S, .sup.32P,
.sup.90Y, .sup.13N, .sup.15O, .sup.211At, .sup.186Re, .sup.188Re,
.sup.75Se), and enzymatic labels (e.g., horseradish peroxidase,
alkaline phosphatase, glucose oxidase, urease,
acetylcholinetransferase, luciferase, and beta-galactosidase),
epitope tags, chromophore labels, phosphorescent labels,
photoaffinity molecules, ECL labels, dyes, biotin, haptens, and the
like. Such labels are well known in the art and can be attached to
the anti-GITR antibodies using art-recognized methods. Preferably,
attachment of the detectable label does not significantly interfere
with binding of the antibody to GITR. The detection means is
determined by the chosen label. Appearance of the label or its
reaction products can be achieved using the naked eye, in the case
where the label is particulate and accumulates at appropriate
levels, or using instruments such as a spectrophotometer, a
luminometer, a fluorimeter, and the like, all in accordance with
standard practice.
Preferably, methods of conjugating the labels result in linkages
which are substantially (or nearly) non-immunogenic, e.g., peptide-
(i.e. amide-), sulfide-, (sterically hindered), disulfide-,
hydrazone-, and ether linkages. These linkages are nearly
non-immunogenic and show reasonable stability within serum (see
e.g. Senter et al., Curr Opin Chem Biol 2009; 13 235-44; WO
2009/059278; WO 95/17886).
Depending on the biochemical nature of the moiety and the antibody,
different conjugation strategies can be employed. In case the
moiety is naturally occurring or recombinant of between 50 to 500
amino acids, there are standard procedures in text books describing
the chemistry for synthesis of protein conjugates, which can be
easily followed by the skilled artisan (see e.g. Hackenberger et
al., Angew Chem Int Ed Engl 2008; 47:10030-74). In one embodiment
the reaction of a maleinimido moiety with a cysteine residue within
the antibody or the moiety is used. This is an especially suited
coupling chemistry in case e.g. a Fab or Fab'-fragment of an
antibody is used. Alternatively in one embodiment coupling to the
C-terminal end of the antibody or moiety is performed. C-terminal
modification of a protein, e.g. of a Fab-fragment can e.g. be
performed as described (Sunbul et al., Org Biomol Chem 2009;
7:3361-71.
In general, site specific reaction and covalent coupling is based
on transforming a natural amino acid into an amino acid with a
reactivity which is orthogonal to the reactivity of the other
functional groups present. For example, a specific cysteine within
a rare sequence context can be enzymatically converted in an
aldehyde (see Frese et al. ChemBioChem 2009; 10:425-7). It is also
possible to obtain a desired amino acid modification by utilizing
the specific enzymatic reactivity of certain enzymes with a natural
amino acid in a given sequence context (see, e.g., Taki et al.,
Prot Eng Des Sel 2004; 17:119-26; Gautier et al. Chem Biol 2008;
15:128-36; Bordusa et al., Bioorganic Chemistry (2004)
389-403).
Site specific reaction and covalent coupling can also be achieved
by the selective reaction of terminal amino acids with appropriate
modifying reagents.
The reactivity of an N-terminal cysteine with benzonitrils (see Ren
et al., Angew Chem Int Ed Engl 2009; 48:9658-62) can be used to
achieve a site-specific covalent coupling.
Native chemical ligation can also rely on C-terminal cysteine
residues (Taylor et al., Nucleic Acids and Molecular Biology 2009;
22:65-96). EP 1 074 563 describes a conjugation method which is
based on the faster reaction of a cysteine within a stretch of
negatively charged amino acids with a cysteine located in a stretch
of positively charged amino acids.
The moiety may also be a synthetic peptide or peptide mimic. In
case a polypeptide is chemically synthesized, amino acids with
orthogonal chemical reactivity can be incorporated during such
synthesis (see, e.g., de Graaf et al., Bioconjug Chem 2009;
20:1281-95). Since a great variety of orthogonal functional groups
is at stake and can be introduced into a synthetic peptide,
conjugation of such peptide to a linker is standard chemistry.
In some embodiments, the detectable label is indirectly conjugated
to the antibody. For example, the antibody can be conjugated to
biotin, and the label can be conjugated to avidin using methods
well known in the art. In another embodiment, an unlabeled antibody
is detected using a labeled antibody which binds to the unlabeled
antibody. In certain embodiments, the labeled antibody that is used
for detection is labeled with any of the labels set forth above. In
certain embodiments, the labeling and detection is conducted as
described in the Examples, e.g., using the Leica Bond Polymer
Refine Detection System (Leica, Buffalo Grove, Ill., Catalog
DS9800).
Methods of Production
The diagnostic anti-GITR antibodies described herein can be
produced using art-recognized techniques. In certain embodiments,
the antibody can be purified from a hybridoma producing the
antibody (e.g., the 6G10 antibody). For example, a hybridoma
producing the antibody can be grown in two-liter spinner-flasks for
monoclonal antibody purification. Supernatants can be filtered and
concentrated before affinity chromatography with protein
A-sepharose (Pharmacia, Piscataway, N.J.). Eluted IgG can be
checked by gel electrophoresis and high performance liquid
chromatography to ensure purity. The buffer solution can be
exchanged into PBS, and the concentration can be determined by
OD280 using the 1.43 extinction coefficient.
In certain embodiments, the antibodies described herein can be
produced recombinantly. For example, DNAs encoding partial or
full-length light and heavy chains described herein can be inserted
into expression vectors such that the genes are operatively linked
to transcriptional and translational control sequences. In this
context, the term "operatively linked" is intended to mean that an
antibody gene is ligated into a vector such that transcriptional
and translational control sequences within the vector serve their
intended function of regulating the transcription and translation
of the antibody gene. The expression vector and expression control
sequences are chosen to be compatible with the expression host cell
used. The antibody light chain gene and the antibody heavy chain
gene can be inserted into separate vector or both genes are
inserted into the same expression vector. The antibody genes are
inserted into the expression vector(s) by standard methods (e.g.,
ligation of complementary restriction sites on the antibody gene
fragment and vector, or blunt end ligation if no restriction sites
are present). The light and heavy chain variable regions of the
antibodies described herein can be used to create full-length
antibody genes of any antibody isotype by inserting them into
expression vectors already encoding heavy chain constant and light
chain constant regions of the desired isotype such that the V.sub.H
segment is operatively linked to the C.sub.H segment(s) within the
vector and the V.sub.L segment is operatively linked to the C.sub.L
segment within the vector.
In addition to the antibody chain genes, recombinant expression
vectors may carry regulatory sequences that control the expression
of the antibody chain genes in a host cell. The term "regulatory
sequence" is intended to include promoters, enhancers and other
expression control elements (e.g., polyadenylation signals) that
control the transcription or translation of the antibody chain
genes. Such regulatory sequences are described, for example, in
Goeddel (Gene Expression Technology. Methods in Enzymology 185,
Academic Press, San Diego, Calif. (1990)). It will be appreciated
by those skilled in the art that the design of the expression
vector, including the selection of regulatory sequences, may depend
on such factors as the choice of the host cell to be transformed,
the level of expression of protein desired, etc. Preferred
regulatory sequences for mammalian host cell expression include
viral elements that direct high levels of protein expression in
mammalian cells, such as promoters and/or enhancers derived from
cytomegalovirus (CMV), Simian Virus 40 (SV40), adenovirus, (e.g.,
the adenovirus major late promoter (AdMLP) and polyoma.
Alternatively, nonviral regulatory sequences may be used, such as
the ubiquitin promoter or .beta.-globin promoter. Still further,
regulatory elements composed of sequences from different sources,
such as the SR.alpha. promoter system, which contains sequences
from the SV40 early promoter and the long terminal repeat of human
T cell leukemia virus type 1 (Takebe et al., MCB 1988;
8:466-72).
In addition to the antibody chain genes and regulatory sequences,
recombinant expression vectors may carry additional sequences, such
as sequences that regulate replication of the vector in host cells
(e.g., origins of replication) and selectable marker genes. The
selectable marker gene facilitates selection of host cells into
which the vector has been introduced (see, e.g., U.S. Pat. Nos.
4,399,216, 4,634,665 and 5,179,017). For example, typically the
selectable marker gene confers resistance to drugs, such as G418,
hygromycin or methotrexate, on a host cell into which the vector
has been introduced. Preferred selectable marker genes include the
dihydrofolate reductase (DHFR) gene (for use in dhfr-host cells
with methotrexate selection/amplification) and the neo gene (for
G418 selection).
For expression of the light and heavy chains, the expression
vector(s) encoding the heavy and light chains is transfected into a
host cell by standard techniques. The various forms of the term
"transfection" are intended to encompass a wide variety of
techniques commonly used for the introduction of exogenous DNA into
a prokaryotic or eukaryotic host cell, e.g., electroporation,
calcium-phosphate precipitation, DEAE-dextran transfection and the
like. Although it is theoretically possible to express the
antibodies described herein in either prokaryotic or eukaryotic
host cells, expression of antibodies in eukaryotic cells, and most
preferably mammalian host cells, is the most preferred because such
eukaryotic cells, and in particular mammalian cells, are more
likely than prokaryotic cells to assemble and secrete a properly
folded and immunologically active antibody. Prokaryotic expression
of antibody genes has been reported to be ineffective for
production of high yields of active antibody (Boss et al.,
Immunology Today 1985; 6:12-13).
Preferred mammalian host cells for expressing the recombinant
antibodies described herein include Chinese Hamster Ovary (CHO
cells) (including dhfr-CHO cells, described in Urlaub and Chasin,
PNAS 19809; 77:4216-220, used with a DHFR selectable marker, e.g.,
as described in Kaufman et al., Mol Biol 1982; 159:601-21, NSO
myeloma cells, COS cells, and SP2 cells. In particular, for use
with NSO myeloma cells, another preferred expression system is the
GS gene expression system disclosed in WO 87/04462, WO 89/01036 and
EP 338,841. When recombinant expression vectors encoding antibody
genes are introduced into mammalian host cells, the antibodies are
produced by culturing the host cells for a period of time
sufficient to allow for expression of the antibody in the host
cells or, more preferably, secretion of the antibody into the
culture medium in which the host cells are grown. Antibodies can be
recovered from the culture medium using standard protein
purification methods.
The chimeric anti-GITR antibodies described herein can be prepared
based on the sequence of a human monoclonal antibody prepared as
described above. For instance, in one embodiment, DNA encoding the
human heavy and light variable regions of the 6G10 antibody can be
operatively linked to a non-human constant region (e.g., a murine
constant region) using routine recombinant technology.
Methods of Detection of Human GITR on Cells
Provided herein are methods for detecting GITR protein expression
in a biological sample, e.g., a human tumor tissue sample, by
contacting the sample with an anti-GITR antibody, e.g., described
herein, such as an anti-GITR antibody comprising the heavy and
light chain variable region CDR sequences set forth in SEQ ID NOs:
5-7 and 8-10, respectively, heavy and light chain variable region
sequences set forth in SEQ ID NOs: 11 and 12, respectively, or full
length heavy and light chain sequences of antibody 6G10 (a fully
human anti-GITR antibody) set forth in SEQ ID NOs: 13 and 14,
respectively, wherein the antibody or antigen-binding fragment
thereof specifically binds to human GITR, and detecting the binding
of the antibody to GITR in the sample, e.g., by
immunohistochemistry. In certain embodiments, the heavy chain lacks
the C-terminal lysine. In some embodiments, the anti-GITR antibody
is a chimeric antibody comprising the variable regions of antibody
6G10, but non-human constant regions (e.g., mouse constant
regions), e.g., a chimeric 6G10 antibody comprising the heavy and
light chain sequences set forth in SEQ ID NOs: 15 and 16,
respectively. In certain embodiments, the heavy chain of the
chimeric antibody lacks the C-terminal lysine.
In certain embodiments, exemplary biological samples include blood
samples, serum samples, cells, surgically-resected tissue, and
biopsied tissue (e.g. cancer tissue) obtained from the cancer
patient. Biological samples for use in the methods described herein
can be fresh, frozen, or fixed. Biological samples, e.g., tumor
samples, can be obtained from the patient using routine methods,
such as, but not limited to, biopsy, surgical resection, or
aspiration.
The methods used to detect GITR expression in human tissue, e.g.,
tumor tissue, can be qualitative, semi-quantitative, or
quantitative. Art-recognized antibody-based methods for detecting
GITR protein levels in biological samples include, but are not
limited to, enzyme-linked immunosorbent assay (ELISA),
radioimmunoassays, electrochemiluninescence (ECL) assays, surface
plasmon resonance, western blot, immunoprecipitation,
fluorescence-activated cell sorting (FACS), immunofluorescence,
immunohistochemistry, and the like. Methods for detecting GITR
expression in a biological sample may include control samples
(negative and positive controls). For example, a negative control
sample may be a sample containing no GITR protein, and a positive
controls sample is a sample containing GITR protein. Comparing the
results with the negative and positive controls can confirm the
presence or absence of GITR in the biological sample.
In one embodiment, GITR expression in a tumor tissue sample is
assessed by immunohistochemistry. This technique allows detection
of GITR on specific cell types, e.g., Treg cells, e.g., Treg cells
within a tumor, and tumor cells.
General immunohistochemical methods are well-known in the art.
Specific conditions, considerations, and guidance for performing
immunohistochemistry are described in, e.g., Immunohistochemical
Staining Methods 6th Edition 2013, Taylor C R, Rudbeck L, eds.,
Dako North America, available as an Education Guide at
www.dako.com; "Immunohistochemical Staining Method guide").
Immunohistochemical staining may be conducted on fixed tissue
sections or on frozen tissue sections. When using fixed tissue
sections, e.g., formalin fixed paraffin embedded (FFPE) tissue
sections, the procedure generally involves the following steps:
obtaining a tumor tissue sample (e.g., by biopsy), fixation of the
tumor sample; embedding (e.g., in paraffin); sectioning and
mounting; antigen retrieval; incubation with a primary antibody
(e.g., chimeric 6G10 antibody described herein), detection (e.g.,
after amplification of the antigen/antibody complex signal), and
interpretation by a pathologist (e.g., using an art-recognized
scoring system).
Suitable, non-limiting fixatives include, for example,
paraformaldehyde, glutaraldehyde, formaldehyde, acetic acid,
acetone, osmium tetroxide, chromic acid, mercuric chloride, picric
acid, alcohols (e.g., methanol, ethanol), Gendre's fluid, Rossman's
fluid, B5 fixative, Bouin's fluid, Carnoy's fixative, and
methacarn. In a preferred embodiment, the tumor sample is fixed in
formaldehyde (e.g., 10% buffered formalin, which corresponds to 4%
formaldehyde in a buffered solution). Once fixed, the tumor sample
is serially dehydrated using alcohol or xylene, embedded in
paraffin, and cut with a microtome to generate tumor tissue
sections (e.g., with a thickness of about 4-5 .mu.m), which can
then be mounted onto microscope slides (e.g., adhesive-coated glass
slides).
Exemplary embedding materials include paraffin, celloidin, OCT.TM.
compound, agar, plastics, or acrylics. In a preferred embodiment,
the embedding agent is paraffin.
Antigen retrieval can be performed using any method known in the
art. Antigen retrieval treatment can involve heat (e.g.,
heat-induced epitope retrieval (HIER), as described in Example 3),
or enzymatic degradation (proteolytic-induced epitope retrieval
(PIER)). Exemplary antigen retrieval methods include microwave
heating, proteolytic treatment (e.g., with proteinase K), pressure
cooker heating, and water bath heating. Detailed general methods
for antigen retrieval are described in Chapter 3 of the
Immunohistochemical Staining Method guide. In certain embodiments,
the antigen retrieval is performed by HIER in combination with
commercially available antigen retrieval buffers (as described in
Example 3).
The methods described herein employ anti-GITR antibodies to detect
the expression levels of human GITR protein. In certain
embodiments, the diagnostic anti-GITR antibody comprises the heavy
and light chain variable region CDR sequences set forth in SEQ ID
NOs: 5-7 and 8-10, respectively, heavy and light chain variable
region sequences set forth in SEQ ID NOs: 11 and 12, respectively,
or full length heavy and light chain sequences of antibody 6G10 (a
fully human anti-GITR antibody) set forth in SEQ ID NOs: 13 and 14,
respectively, wherein the antibody or antigen-binding fragment
thereof specifically binds to human GITR, as described supra. In
certain embodiments, the heavy chain lacks the C-terminal lysine.
In some embodiments, the anti-GITR antibody is a chimeric antibody
comprising the variable regions of antibody 6G10, but non-human
constant regions (e.g., mouse constant regions), e.g., a chimeric
6G10 antibody comprising the heavy and light chain sequences set
forth in SEQ ID NOs: 15 and 16, respectively. In certain
embodiments, the heavy chain of the chimeric antibody lacks the
C-terminal lysine. Such antibodies are particularly useful for
detecting GITR expression in tumor tissue sections (see, e.g.,
Example 3).
GITR protein expression in tissue, e.g., tumor, samples can be
detected using direct or indirect methods. For example, the primary
antibody may comprise a detectable moiety, such as the enzyme
horseradish peroxidase (HRP) or a fluorescent label (e.g., FITC,
TRITC), as described herein. In certain embodiments, the primary
antibody does not itself comprise a detectable moiety, but is
detected by binding of a secondary antibody to it, for indirect
immunohistochemistry. Accordingly, in certain embodiments, the
secondary antibody comprises a detectable label, e.g., an
enzymatic, chromogenic, or fluorescent label. In certain
embodiments, the primary antibody is a chimeric antibody comprising
human variable region sequences and a non-human Fc region. A
secondary antibody can be used to recognize the non-human Fc region
of the primary antibody in order to reduce background staining.
In some embodiments, GITR protein expression can be detected using
the avidin-biotin complex method (ABC method). In these
embodiments, the secondary antibody is biotinylated and can serve
as a bridge between the primary antibody and
biotin-avidin-peroxidase complex. Other suitable non-limiting
methods for immunohistochemistry include those described in Chapter
6 of the Immunohistochemical Staining Method guide, e.g., methods
described in Chilosi et al., Biotech Histochem 1994; 69:235;
Sabattini et al., J Clin Pathol 1998; 51:506-11; and Gross et al.,
JBC 1959; 234:1622.
General methods for preparing and staining frozen tissue sections
are well known in the art, and are described in, e.g., the
Immunohistochemical Staining Method guide, Chapter 3 of
Immunohistochemistry and Methods (Buchwalow and Bocker,
Springer-Verlag Berlin Heidelberg 2010), and Chapter 21 of Theory
and Practice of Histological Techniques (Bancroft and Gamble,
6.sup.th Edition, Elsevier Ltd., 2008). Frozen tissue section
staining can be performed as described in Example 3.
GITR protein can be detected as described in the Examples.
In some embodiments, slides stained with the anti-GITR antibodies
described herein can be further counterstained with, e.g.,
hematoxylin and/or eosin, using methods well known in the art.
In the interpretation step, GITR protein expression detected by
immunohistochemistry can be scored using art-recognized scoring
methods. Non-limiting examples of scoring methods are described
below.
For example, in certain embodiments, GITR expression is evaluated
using the H-score (histochemical score) system, which is widely
used in the art and is useful given its dynamic range and use of
weighted percentiles. The H-score is a semiquantitative scoring
system based on the formula: 3.times.percentage of strongly
staining cells (3+ staining)+2.times.percentage of moderately
staining (2+ staining) cells+1.times.percentage of weakly staining
(1+ staining) cells+0.times.percentage of non-stained (0 staining)
cells, giving a score range of 0 to 300. See, e.g., McCarty et al.,
Cancer Res 1986; 46:4244-8; Bosman et al., J Clin Pathol 1992;
45:120-4; Dieset et al., Analyt Quant Cytol Histol 1996; 18:351-4.
Control tissue can include, e.g., matching non-tumor tissue from
the same subject. Accordingly, in certain embodiments, the number
of cells staining positive for GITR using the anti-GITR antibodies
herein, and the intensity of staining, can be used to determine an
H-score.
In other embodiments, GITR expression is evaluated using the Allred
scoring system (Allred et al., Mod Pathol 1998; 11:155-68; Harvey
et al., J Clin Oncol 1999; 17:1474-91). This scoring system
involves adding proportion and intensity scores to obtain a total
score. Accordingly, in some embodiments, the proportion score is
obtained based on the estimated proportion of tumor cells that are
positive for GITR (0: none, 1: < 1/100, 2: 1/100 to 1/10, 3:
1/10 to 1/3; 4: 1/3 to 2/3; and 5: >2/3), and the intensity
score is obtained based on the average intensity of GITR expression
in positive tumor cells (0: none, 1: weak, 2: intermediate, 3:
strong). Thus, Allred scores range from 0 to 8, with a score
ranging from 3 to 8 considered positive (i.e., positive detection).
Accordingly, in certain embodiments, tumors with an Allred score of
3 to 8, for example, an Allred score of 3, 4, 5, 6, 7, or 8, for
GITR staining are considered GITR-positive tumors.
In other embodiments, the scoring system is automated, e.g.,
computerized, and quantified by image analysis. Automated methods
are well known in the art. For example, an average threshold
measure (ATM) score, which obtains an average of 255 staining
intensity levels, can be calculated as described in, e.g.,
Choudhury et al., J Histochem Cytochem 2010; 58:95-107, Rizzardi et
al., Diagnostic Pathology 2012; 7:42-52. Another automated scoring
system is AQUA.RTM. (automated quantitative analysis), which is
performed using, e.g., tissue microarrays (TMAs), on a continuous
scale. AQUA.RTM. is a hybrid of standard immunohistochemistry and
flow cytometry that provides an objective numeric score ranging
from 1-255, and involves antigen retrieval, use of primary and
secondary antibodies, and multiplexed fluorescent detection.
Optimal cutoff points can be determined as described in Camp et al.
Clin Cancer Res 2004; 10:7252-9. The AQUA.RTM. scoring system is
described in detail in Camp et al., Nat Med 2002; 8:1323-7; Camp et
al., Cancer Res 2003; 63:1445-8; Ghosh et al., Hum Pathol 2008;
39:1835-43; Bose et al., BMC Cancer 2012; 12:332; Mascaux et al.,
Clin Cancer Res 2011; 17:7796-807. Other suitable automated
immunohistochemistry platforms include commercially available
platforms, such as the Leica BOND RX staining platform. The
platform detects GITR expression in tumor tissue on the basis of
staining intensity on the following scale: minimal, <1 cells per
20.times. objective field; mild, 1.about.<10 cells per 20.times.
objective field; moderate, 10.about.<50 cells per 20.times.
objective field, marked, 50.about.<200 cells per 20.times.
objective; and intense, >200 cells per 20.times. objective
field.
Accordingly, provided herein are methods for determining whether a
tumor of a cancer patient is GITR-positive and/or comprises GITR
positive cells, comprising contacting a tumor sample from the
patient with an antibody, or antigen-binding portion thereof,
comprising heavy chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 5-7,
respectively, and light chain variable region CDR1, CDR2, and CDR3
comprising the amino acid sequences set forth in SEQ ID NOs: 8-10,
respectively, wherein the antibody or antigen-binding fragment
thereof specifically binds to human GITR.
Also provided herein are methods for determining whether a tumor of
a cancer patient is GITR-positive and/or comprises GITR positive
cells, comprising:
(a) contacting a tumor sample from the patient with an antibody, or
antigen-binding portion thereof, comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human GITR,
and
(b) detecting the binding of the antibody to GITR in the
sample.
Also provided herein are methods for determining whether a tumor of
a cancer patient is GITR-positive and/or comprises GITR positive
cells comprising:
(a) contacting a tumor sample from the patient with an antibody, or
antigen-binding portion thereof, comprising heavy chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 5-7, respectively, and light chain variable
region CDR1, CDR2, and CDR3 comprising the amino acid sequences set
forth in SEQ ID NOs: 8-10, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human
GITR,
(b) detecting the binding of the antibody to GITR in the sample,
and
(c) determining the level of GITR protein expression in the sample,
wherein a level of GITR protein above a threshold level, e.g.,
level of detection using a given detection method, indicates that
the tumor is GITR-positive tumor.
Also provided herein are methods for identifying GITR positive
cells in a tissue (e.g., tumor) sample comprising:
(a) contacting a tissue (e.g., tumor) sample from the patient with
an antibody, or antigen-binding portion thereof, comprising heavy
chain variable region CDR1, CDR2, and CDR3 comprising the amino
acid sequences set forth in SEQ ID NOs: 5-7, respectively, and
light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on specific or
individual cells of the tissue (e.g., tumor) sample; and
optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
given cell types (e.g., CD3, CD4, CD8, CD25, or FoxP3 markers)
and/or with hematoxylin and eosin, to identify the cells that were
identified as GITR positive in step (b).
Step (c) allows identification of GITR positive cells as being,
e.g., T cells, such as Treg cells, or tumor cells. Thus, provided
herein are the following methods:
A method for detecting the presence of, and/or identifying the
number of, T cells expressing GITR in a tissue (e.g., tumor)
sample, and/or level of GITR on T cells of a tissue (e.g., tumor)
sample comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on T cells of the
tissue (e.g., tumor) sample; and optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
T cells (e.g., CD3, CD4, CD8, CD25, or FoxP3 markers) and/or with
hematoxylin and eosin, to identify the T cells that were identified
as GITR positive in step (b).
A method for detecting the presence of, and/or identifying the
number of, Treg cells expressing GITR in a tissue (e.g., tumor)
sample, and/or the level of GITR on Treg cells of a tissue (e.g.,
tumor) sample comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on Treg cells of
the tissue (e.g., tumor) sample; and optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
Treg cells (e.g., a FoxP3 marker) and/or with hematoxylin and
eosin, to identify the Treg cells that were identified as GITR
positive in step (b).
A method for detecting the presence of, and/or identifying the
number of, Teff cells expressing GITR in a tissue (e.g., tumor)
sample, and/or the level of GITR on Teff cells of a tissue (e.g.,
tumor) sample comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on Teff cells of
the tissue (e.g., tumor) sample; and optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
Teff cells (e.g., a FoxP3 marker) and/or with hematoxylin and
eosin, to identify the Teff cells that were identified as GITR
positive in step (b).
A method for detecting the presence of, and/or identifying the
number of Treg and Teff cells expressing GITR in a tissue (e.g.,
tumor) sample, and/or the level of GITR on Treg and Teff cells of a
tissue (e.g., tumor) sample comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on Treg and Teff
cells of the tissue (e.g., tumor) sample; and optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
Treg and Teff cells (e.g., CD3, CD4, CD8, CD25, and/or FoxP3
markers) and/or with hematoxylin and eosin, to identify the Treg
and Teff cells that were identified as GITR positive in step
(b).
A method for detecting the presence of, or identifying the number
of, tumor cells expressing GITR in a tumor sample, and/or level of
GITR on tumor cells of a tumor sample comprising:
(a) contacting a tumor sample, e.g., from a subject, with an
antibody, or antigen-binding portion thereof, comprising heavy
chain variable region CDR1, CDR2, and CDR3 comprising the amino
acid sequences set forth in SEQ ID NOs: 5-7, respectively, and
light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on tumor cells of
the tumor sample; and optionally
(c) staining the tumor sample, e.g., with markers of tumor cells or
other cell type and/or with hematoxylin and eosin, to identify the
tumor cells that were identified as GITR positive in step (b).
A method for detecting the presence of, and/or identifying the
number of, tumor cells and Treg cells expressing GITR in a tissue
(e.g., tumor) sample, and/or the level of GITR on tumor cells and
Treg cells of a tissue (e.g., tumor) sample comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on tumor cells
and Treg cells of the tissue (e.g., tumor) sample; and
optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
tumor and/or Treg cells (e.g., a FoxP3 marker) and/or with
hematoxylin and eosin, to identify the tumor and Treg cells that
were identified as GITR positive in step (b).
A method for detecting the presence of, and/or identifying the
number of, tumor cells, Treg cells and Teff cells expressing GITR
in a tissue (e.g., tumor) sample, and/or the level of GITR on tumor
cells, Treg cells and Teff cells of a tissue (e.g., tumor) sample
comprising:
(a) contacting a tissue (e.g., tumor) sample, e.g., from a subject,
with an antibody, or antigen-binding portion thereof, comprising
heavy chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 5-7, respectively,
and light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR; and
(b) detecting the binding of the antibody to GITR on tumor cells,
Treg cells and Teff cells of the tissue (e.g., tumor) sample; and
optionally
(c) staining the tissue (e.g., tumor) sample, e.g., with markers of
tumor cells, Treg cells and/or Teff cells (e.g., CD3, CD4, CD8,
CD25, and/or FoxP3 markers) and/or with hematoxylin and eosin, to
identify the tumor cells, Treg cells and Teff cells that were
identified as GITR positive in step (b).
Exemplary tumor markers include, for example, ErbB receptors, Melan
A [MART1], gp100, tyrosinase, TRP-1/gp 75, TRP-2, MAGE-1, MAGE-3,
HPV E6 and E7 proteins, Mucin [MUC-1], prostate-specific antigen
[PSA], carcinoembryonic antigen [CEA] PIA tumor antigen, MAGE-2,
MAGE-4, MAGE-6, MAGE-10, MAGE-12, BAGE-1, CAGE-1, 2, 8, CAGE-3 to
7, LAGE-1, NY-ESO-1/LAGE-2, NA-88, GnTV, and TRP2-INT2.
T reg cell surface markers include CD4, CD25 and FOXP3.
In certain embodiments, a combination of detection methods is used
to increase the accuracy of designating a tumor as being
GITR-positive or GITR-negative. For example, in one embodiment, a
combination of immunohistochemistry and fluorescence in situ
hybridization (FISH) is used to type a tumor as being GITR positive
or negative. Such approaches have been used to, e.g., optimize
Her-2/neu testing. See, e.g., Ridolfi et al., Mod Pathol 2000;
13:866-73. Any of the antibody-based methods of detection described
herein can be used in the combination, or in combination with
RNA-based detection methods, such as RNAscope and quantitative
PCR.
The methods of detection described above are typically performed in
conjunction with a reference or control sample. In some
embodiments, the control sample is a section of matched normal
tissue. In some embodiments, the control or reference sample is
derived from a subject that is not afflicted with a GITR-expressing
tumor or cancer (e.g., a healthy subject). Such samples may provide
a standardized reference for the amount of GITR-expressing cells in
a biological sample.
Exemplary Prognostic Methods
Provided herein are methods for identifying a cancer patient that
is likely to respond to an anti-GITR immunotherapy or predicting
whether a cancer patient will respond to anti-GITR immunotherapy
comprising:
(a) contacting a tumor sample from the patient with an anti-GITR
antibody, or antigen-binding portion thereof,
(b) detecting the binding of the antibody to GITR in the tumor
sample,
(c) determining the level of GITR protein expression in the sample,
wherein a level of GITR protein above a threshold level indicates
that the tumor is a GITR-positive tumor and that the patient is
likely to or predicted to respond to anti-GITR immunotherapy. In
certain embodiments, the level of expression of GITR on specific
immune cells, e.g., tumor infiltration lymphocytes (TILs), such as
T cells, e.g., Treg cells, e.g., in a tumor, is detected or
monitored, and, e.g., determines likelihood of response of a
subject to an anti-GITR therapy. In certain embodiments, the levels
of expression of GITR on tumor cells is is detected or monitored,
and, e.g., determines likelihood of response of a subject to an
anti-GITR therapy.
In certain embodiments, the anti-GITR antibody is an antibody
comprising the heavy and light chain variable region CDR sequences
set forth in SEQ ID NOs: 5-7 and 8-10, respectively, heavy and
light chain variable region sequences set forth in SEQ ID NOs: 11
and 12, respectively, or full length heavy and light chain
sequences of antibody 6G10 (a fully human anti-GITR antibody) set
forth in SEQ ID NOs: 13 and 14, wherein the antibody or
antigen-binding fragment thereof specifically binds to human GITR.
In certain embodiments, the heavy chain lacks the C-terminal
lysine. In some embodiments, the anti-GITR antibody is a chimeric
antibody comprising the variable regions of antibody 6G10, but
non-human constant regions (e.g., mouse constant regions), e.g., a
chimeric 6G10 antibody comprising the heavy and light chain
sequences set forth in SEQ ID NOs: 15 and 16, respectively. In
certain embodiments, the heavy chain of the chimeric antibody lacks
the C-terminal lysine. In certain embodiments, the detection of
GITR expression is performed by immunohistochemistry, as described
supra. In one embodiment, immunohistochemistry is combined with
another detection method, e.g., FISH.
In certain embodiments, the tumor sample is prepared as FFPE
sections. In other embodiments, the tumor sample is prepared as
frozen tissue sections.
In some embodiments, a tumor is considered GITR-positive and likely
to benefit from anti-GITR immunotherapy if a sample of the tumor or
immune cells therein, e.g., tumor infiltration lymphocytes (TILs),
e.g., Treg cells, has more GITR-positive cells than a matching
normal tissue, or immune cell, e.g., T reg cell, from the same
cancer patient.
In certain embodiments, the cancer patient is likely to or
predicted to benefit from anti-GITR immunotherapy if the number of
GITR-positive cells, e.g., immune cells, e.g., tumor infiltration
lymphocytes (TILs), such as T reg cells, and/or tumor cells, in the
tumor sample exceeds a threshold level. A threshold level may be
the lowest level of GITR as determined with a given detection
system, e.g., immunohistochemistry, as further described herein,
such as in the Examples. In certain embodiments, the cancer patient
is likely to or predicted to benefit from anti-GITR immunotherapy
if the number of GITR-positive cells, e.g., immune cells, e.g.,
tumor infiltration lymphocytes (TILs), such as T reg cells, and/or
tumor cells, in the tumor sample exceeds a threshold number or
proportion using an art-recognized scoring system described supra.
Thus, in some embodiments, the number or proportion of
GITR-positive cells exceeds a certain threshold number or
proportion using a scoring system described herein and is defined
as being "GITR-positive."
In certain embodiments, a tumor sample having at least 1%, for
example, at least 5%, at least 10%, at least 15%, at least 20%, at
least 25%, at least 30%, at least 35%, at least 40%, at least 45%,
at least 50%, at least 55%, at least 60%, at least 65%, at least
70%, at least 75%, at least 80%, at least 90%, or at least 95% of
cells in the tumor, e.g., tumor cells and/or immune cells, e.g.,
tumor infiltration lymphocytes (TILs), such as T reg cells,
expressing GITR protein is designated as being "GITR-positive" and
indicates that the cancer patient is likely to or predicted to
respond to anti-GITR immunotherapy.
In some embodiments, an H score is calculated for the tumor sample.
Accordingly, in certain embodiments, a tumor sample with an H score
of at least 5, such as at least 10, at least 15, at least 20, at
least 25, at least 30, at least 40, at least 50, at least 60, at
least 70, at least 80, at least 90, at least 100, at least 125, at
least 150, at least 175, at least 200, at least 225, at least 250,
at least 275, or at least 290 is designated as being
"GITR-positive" and indicates that the cancer patient is likely to
or predicted to respond to anti-GITR immunotherapy. The score may
be calculated based on all cells in a tumor, or a specific type of
cells, e.g., immune cells, e.g., tumor infiltration lymphocytes
(TILs), such as Treg cells, and/or tumor cells.
In some embodiments, an Allred score is calculated for the tumor
sample. Accordingly, in certain embodiments, a tumor sample with an
Allred score of at least 3, such as at least 4, at least 5, at
least 6, at least 7, or 8 is designated as being "GITR-positive"
and indicates that the cancer patient is likely to or predicted to
respond to anti-GITR immunotherapy. The score may be calculated
based on all cells in a tumor, or a specific type of cells, e.g.,
immune cells, e.g., tumor infiltration lymphocytes (TILs), such as
Treg cells, and/or tumor cells.
In some embodiments, an AQUA score is calculated. Accordingly, in
certain embodiments, a tumor sample with an AQUA score of at least
5, such as at least 10, at least 15, at least 20, at least 25, at
least 30, at least 40, at least 50, at least 60, at least 70, at
least 80, at least 90, at least 100, at least 125, at least 150, at
least 175, at least 200, at least 225, or at least 250 is
designated as being "GITR-positive" and indicates that the cancer
patient is likely to or predicted to respond to anti-GITR
immunotherapy. The score may be calculated based on all cells in a
tumor, or a specific type of cells, e.g., immune cells, e.g., tumor
infiltration lymphocytes (TILs), such as Treg cells, and/or tumor
cells.
In certain embodiments, a combination of one of the scoring systems
above with another detection modality, such as FISH, is used to
increase the accuracy of determining whether a tumor is
GITR-positive or GITR-negative.
In certain embodiments, the patient has a cancer selected from the
group consisting of squamous cell carcinoma, small-cell lung
cancer, non-small cell lung cancer, squamous non-small cell lung
cancer (NSCLC), non squamous NSCLC, glioma, gastrointestinal
cancer, renal cancer (e.g. clear cell carcinoma), ovarian cancer,
liver cancer, colorectal cancer, endometrial cancer, kidney cancer
(e.g., renal cell carcinoma (RCC)), prostate cancer (e.g. hormone
refractory prostate adenocarcinoma), thyroid cancer, neuroblastoma,
pancreatic cancer, glioblastoma (glioblastoma multiforme), cervical
cancer, stomach cancer, bladder cancer, hepatoma, breast cancer,
colon carcinoma, and head and neck cancer (or carcinoma), gastric
cancer, germ cell tumor, pediatric sarcoma, sinonasal natural
killer, melanoma (e.g., metastatic malignant melanoma, such as
cutaneous or intraocular malignant melanoma), bone cancer, skin
cancer, uterine cancer, cancer of the anal region, testicular
cancer, carcinoma of the fallopian tubes, carcinoma of the
endometrium, carcinoma of the cervix, carcinoma of the vagina,
carcinoma of the vulva, cancer of the esophagus, cancer of the
small intestine, cancer of the endocrine system, cancer of the
parathyroid gland, cancer of the adrenal gland, sarcoma of soft
tissue, cancer of the urethra, cancer of the penis, solid tumors of
childhood, cancer of the ureter, carcinoma of the renal pelvis,
neoplasm of the central nervous system (CNS), primary CNS lymphoma,
tumor angiogenesis, spinal axis tumor, brain cancer, brain stem
glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer,
squamous cell cancer, T-cell lymphoma, environmentally-induced
cancers including those induced by asbestos, virus-related cancers
or cancers of viral origin (e.g., human papilloma virus
(HPV-related or -originating tumors)), and hematologic malignancies
derived from either of the two major blood cell lineages, i.e., the
myeloid cell line (which produces granulocytes, erythrocytes,
thrombocytes, macrophages and mast cells) or lymphoid cell line
(which produces B, T, NK and plasma cells), such as all types of
leukemias, lymphomas, and myelomas, e.g., acute, chronic,
lymphocytic and/or myelogenous leukemias, such as acute leukemia
(ALL), acute myelogenous leukemia (AML), chronic lymphocytic
leukemia (CLL), and chronic myelogenous leukemia (CML),
undifferentiated AML (M0), myeloblastic leukemia (M1), myeloblastic
leukemia (M2; with cell maturation), promyelocytic leukemia (M3 or
M3 variant [M3V]), myelomonocytic leukemia (M4 or M4 variant with
eosinophilia [M4E]), monocytic leukemia (M5), erythroleukemia (M6),
megakaryoblastic leukemia (M7), isolated granulocytic sarcoma, and
chloroma; lymphomas, such as Hodgkin's lymphoma (HL), non-Hodgkin's
lymphoma (NHL), B cell hematologic malignancy, e.g., B-cell
lymphomas, T-cell lymphomas, lymphoplasmacytoid lymphoma,
monocytoid B-cell lymphoma, mucosa-associated lymphoid tissue
(MALT) lymphoma, anaplastic (e.g., Ki 1+) large-cell lymphoma,
adult T-cell lymphoma/leukemia, mantle cell lymphoma, angio
immunoblastic T-cell lymphoma, angiocentric lymphoma, intestinal
T-cell lymphoma, primary mediastinal B-cell lymphoma, precursor
T-lymphoblastic lymphoma, T-lymphoblastic; and lymphoma/leukaemia
(T-Lbly/T-ALL), peripheral T-cell lymphoma, lymphoblastic lymphoma,
post-transplantation lymphoproliferative disorder, true histiocytic
lymphoma, primary central nervous system lymphoma, primary effusion
lymphoma, B cell lymphoma, lymphoblastic lymphoma (LBL),
hematopoietic tumors of lymphoid lineage, acute lymphoblastic
leukemia, diffuse large B-cell lymphoma, Burkitt's lymphoma,
follicular lymphoma, diffuse histiocytic lymphoma (DHL),
immunoblastic large cell lymphoma, precursor B-lymphoblastic
lymphoma, cutaneous T-cell lymphoma (CTLC) (also called mycosis
fungoides or Sezary syndrome), and lymphoplasmacytoid lymphoma
(LPL) with Waldenstrom's macroglobulinemia; myelomas, such as IgG
myeloma, light chain myeloma, nonsecretory myeloma, smoldering
myeloma (also called indolent myeloma), solitary plasmocytoma, and
multiple myelomas, chronic lymphocytic leukemia (CLL), hairy cell
lymphoma; hematopoietic tumors of myeloid lineage, tumors of
mesenchymal origin, including fibrosarcoma and rhabdomyoscarcoma;
seminoma, teratocarcinoma, tumors of the central and peripheral
nervous, including astrocytoma, schwannomas; tumors of mesenchymal
origin, including fibrosarcoma, rhabdomyoscaroma, and osteosarcoma;
and other tumors, including melanoma, xeroderma pigmentosum,
keratoacanthoma, seminoma, thyroid follicular cancer and
teratocarcinoma, hematopoietic tumors of lymphoid lineage, for
example T-cell and B-cell tumors, including but not limited to
T-cell disorders such as T-prolymphocytic leukemia (T-PLL),
including of the small cell and cerebriform cell type; large
granular lymphocyte leukemia (LGL) preferably of the T-cell type;
a/d T-NHL hepatosplenic lymphoma; peripheral/post-thymic T cell
lymphoma (pleomorphic and immunoblastic subtypes); angiocentric
(nasal) T-cell lymphoma; cancer of the head or neck, renal cancer,
rectal cancer, cancer of the thyroid gland; acute myeloid lymphoma,
as well as any combinations of said cancers. In certain
embodiments, the cancer is a metastatic cancer, refractory cancer,
or recurrent cancer.
Once a cancer patient has been identified as being likely to
benefit from anti-GITR immunotherapy (i.e., has a GITR-positive
tumor), the patient can be treated with a therapeutic anti-GITR
antibody (i.e., anti-GITR immunotherapy).
Accordingly, provided herein are method of treating a cancer
patient with a GITR-positive tumor, as determined by the level of
GITR protein expression using the anti-GITR antibodies and methods
provided herein, comprising administering to the patient a
therapeutically-effective amount of an anti-GITR antibody, or
antigen-binding portion thereof.
In certain embodiments, the anti-GITR antibody used to determine
the level of GITR protein expression is an antibody comprising the
heavy and light chain variable region CDR sequences set forth in
SEQ ID NOs: 5-7 and 8-10, respectively, heavy and light chain
variable region sequences set forth in SEQ ID NOs: 11 and 12,
respectively, or full length heavy and light chain sequences of
antibody 6G10 (a fully human anti-GITR antibody) set forth in SEQ
ID NOs: 13 and 14, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human GITR.
In certain embodiments, the heavy chain lacks the C-terminal
lysine. In some embodiments, the antibody is a chimeric antibody
comprising the variable regions of antibody 6G10, but non-human
constant regions (e.g., mouse constant regions), e.g., a chimeric
6G10 antibody comprising the heavy and light chain sequences set
forth in SEQ ID NOs: 15 and 16, respectively. In certain
embodiments, the heavy chain of the chimeric antibody lacks the
C-terminal lysine. In certain embodiments, the detection of GITR
expression is performed by immunohistochemistry, as described
supra.
In certain embodiments, a cancer patient determined as having a
GITR-positive tumor (ie., GITR-positive tumor cells and/or immune
cells, e.g., tumor infiltration lymphocytes (TILs), such as T reg
cells) using the methods described herein is treated with any agent
that modulates GITR signaling (e.g., an agonist anti-GITR antibody)
and results in, e.g., the induction and/or enhancement of T cell
activation (e.g., increase in IL-2 and/or IFN-.gamma. production by
T cells and/or increase in proliferation of T cells), depletion of
regulatory T cells, etc. Accordingly, provided herein are methods
of treating a cancer patient comprising:
(a) determining whether a tumor (tumor cells and/or immune cells,
e.g., tumor infiltration lymphocytes (TILs), such as Treg cells) of
a cancer patient is GITR-positive using the methods described
herein, e.g., contacting a tumor sample from the patient with an
antibody, or antigen-binding portion thereof, comprising heavy
chain variable region CDR1, CDR2, and CDR3 comprising the amino
acid sequences set forth in SEQ ID NOs: 5-7, respectively, and
light chain variable region CDR1, CDR2, and CDR3 comprising the
amino acid sequences set forth in SEQ ID NOs: 8-10, respectively,
wherein the antibody or antigen-binding fragment thereof
specifically binds to human GITR, detecting the binding of the
antibody to GITR in the sample, and determining the level of GITR
protein expression in the sample, wherein a level of GITR protein
above a threshold level in tumor cells and/or immune cells therein,
e.g., tumor infiltration lymphocytes (TILs), such as Treg cells,
indicates that the tumor is GITR-positive, and
(b) if the tumor is determined to be GITR-positive, then
administering a therapeutically effective amount of an agent that
modulates GITR signaling, e.g., an agent that increases IL-2,
increases IFN-.gamma. production by T cells, and/or increases the
proliferation of T cells and/or depletion of regulatory T
cells.
For example, in one embodiment, the patient is treated with a
therapeutically-effective amount of an agonist anti-GITR antibody
disclosed in WO15187835, the contents of which are herein
incorporated by reference, e.g., antibodies having the heavy and
light chain variable region CDRs, heavy and light chain variable
regions, or heavy and light chains of antibodies 28F3, 19D3, 18E10,
3C3-1, 3C3-2, 2G6, 9G7-1, 9G7-2, 14E3, 19H8-1, 19H8-2, and/or 6G10,
and variants thereof. The sequences of the antibodies disclosed in
PCT Patent Application No. WO15187835 are provided in Table 2 (see
SEQ ID NOs: 5-14 and 27-228). The patient may also be treated with
any other anti-GITR antibodies, e.g., TRX518 (Leap Therapeutics),
MK-4166 (Merck), LKZ-145 (Novartis), GWN-323 (Novartis
Pharmaceuticals Corp.), Medi 1873 (MedImmune), INBRX-110 (Inhibrx),
GITR-Fc protein (OncoMed) and antibodies described in WO2006105021,
WO2009009116, WO2011028683, US2014/0072565, US20140072566,
US20140065152, WO2015031667, WO15184099, WO2015184099, or
WO2016054638.
Exemplary Methods of Monitoring GITR-Positive Tumors/Cancer
Provided herein are methods for monitoring a GITR-expressing
tumor(s) in a cancer patient. Methods for monitoring may be
performed in a similar manner to methods for determining whether a
cancer patient is likely to respond to anti-GITR imp immunotherapy
described above.
The presence of GITR may be monitored in whole tissues, e.g.,
tumors, or in specific cells thereof. For example, the presence of
GITR may be monitored on tumor cells and/or tumor infiltrating
lymphocytes (TILs), e.g., T reg cells and Teff cells.
Monitoring GITR presence (or expression) may comprise determining
GITR presence at two or more time points. For example, a first and
a second tissue sample may be obtained (and the presence and/or
level of GITR determined) from a subject, wherein, e.g., the first
and the second tissue sample are obtained within 3-7 days, 1 week
to 3 weeks, or 1 month to 3 months from each other. A tissue sample
may also be obtained for determining the presence and/or level of
GITR every week or every month.
Accordingly, in certain embodiments, a method for monitoring a
GITR-expressing tumor in a cancer patient comprises:
(a) detecting GITR protein expression in a tissue (e.g., tumor)
sample at a first time point by using an anti-GITR antibody, or
antigen-binding portion thereof,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in a tissue (e.g., tumor,
which may be the same tumor as the one that was biopsied at the
first time point) sample at a second time point using the same
antibody used in step (a), and
(d) determining the level of GITR protein expression in the tumor
from the second time point.
Also provided is a method for monitoring a GITR-expressing tumor in
a cancer patient comprising:
(a) detecting GITR protein expression in Treg cells in a tissue
(e.g., tumor) sample at a first time point by using an anti-GITR
antibody, or antigen-binding portion thereof,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in Treg cells in a tissue
(e.g., tumor, which may be the same tumor as the one that was
biopsied at the first time point) sample at a second time point
using the same antibody used in step (a),
(d) determining the level of GITR protein expression in the Treg
cells from the second time.
Also provided is a method for monitoring a GITR-expressing tumor in
a cancer patient comprising:
(a) detecting GITR protein expression in Teff cells in a tissue
(e.g., tumor) sample at a first time point by using an anti-GITR
antibody, or antigen-binding portion thereof,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in Teff cells in a tissue
(e.g., tumor, which may be the same tumor as the one that was
biopsied at the first time point) sample at a second time point
using the same antibody used in step (a),
(d) determining the level of GITR protein expression in the Teff
cells from the second time.
Also provided is a method for monitoring a GITR-expressing tumor in
a cancer patient comprising:
(a) detecting GITR protein expression in tumor cells in a tumor
sample at a first time point by using an anti-GITR antibody, or
antigen-binding portion thereof,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in tumor cells in a tumor
sample (which may be from the same tumor as the one that was
biopsied at the first time point) at a second time point using the
same antibody used in step (a),
(d) determining the level of GITR protein expression in the tumor
cells from the second time.
In certain methods, the presence and/or level of GITR are
determined in Treg cells and/or Teff cells and/or tumor cells.
Accordingly, in certain embodiments, a method for monitoring a
GITR-expressing tumor in a cancer patient comprises:
(a) detecting GITR protein expression in the GITR-positive tumor at
a first time point by using an anti-GITR antibody, or
antigen-binding portion thereof,
(b) determining the level of GITR protein expression at the first
time point,
(c) detecting GITR protein expression in the tumor at a second time
point using the same antibody used in step (a),
(d) determining the level of GITR protein expression in the tumor
from the second time point; and
(e) comparing the levels of GITR protein expression determined at
the first and second time points.
GITR expression may be determined in the tumor cells and/or in
immune cells, e.g., tumor infiltration lymphocytes (TILs), such as
T reg cells and/or Teff cells within the tumor. A higher level at
the first time point relative to the second time point may be
indicative of tumor regression, a lower score at the first time
point relative to the second time point may be indicative of tumor
progression, and a relatively unchanged level at the first time
point relative to the second time point may be indicative of a
stable tumor.
In certain embodiments, the anti-GITR antibody used to determine
the level of GITR protein expression is an antibody comprising the
heavy and light chain variable region CDR sequences set forth in
SEQ ID NOs: 5-7 and 8-10, respectively, heavy and light chain
variable region sequences set forth in SEQ ID NOs: 11 and 12,
respectively, or full length heavy and light chain sequences of
antibody 6G10 (a fully human anti-GITR antibody) set forth in SEQ
ID NOs: 13 and 14, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human GITR.
In certain embodiments, the heavy chain lacks the C-terminal
lysine. In some embodiments, the antibody is a chimeric antibody
comprising the variable regions of antibody 6G10, but non-human
constant regions (e.g., mouse constant regions), e.g., a chimeric
6G10 antibody comprising the heavy and light chain sequences set
forth in SEQ ID NOs: 15 and 16, respectively. In certain
embodiments, the heavy chain of the chimeric antibody lacks the
C-terminal lysine. In a preferred embodiment, the detection of GITR
expression is performed by immunohistochemistry, as described
herein. In some embodiments, the level of GITR protein expression
is determined using a scoring system selected from the group
consisting of: the H score system, the Allred score system, an
AQUA.RTM. system, or an automated scoring system, as described
herein.
Such methods of monitoring are also useful for determining whether
or not anti-GITR immunotherapy may be effective.
Accordingly, provided herein are methods for monitoring the
efficacy of anti-GITR immunotherapy in a patient having a
GITR-positive tumor comprising:
(a) detecting GITR protein expression in the GITR-positive tumor at
a first time point before or after initiating anti-GITR
immunotherapy by using an anti-GITR antibody,
(b) determining the level of GITR protein expression in the tumor
from the first time point,
(c) detecting GITR protein expression in the GITR-positive tumor at
a second time point after initiating anti-GITR immunotherapy using
the same antibody from step (a),
(d) determining the level of GITR protein expression in the tumor
from the second time point;
(e) comparing the levels of GITR protein expression determined at
the first and second time points, wherein a higher level at the
first time point relative to the second time point may be
indicative of effective anti-GITR immunotherapy, a lower score at
the first time point relative to the second time point is
indicative of ineffective anti-GITR immunotherapy, and an unchanged
score at the first time point relative to the second time point may
be indicative of anti-GITR immunotherapy being stabilizing. GITR
expression may be determined in the tumor cells and/or in immune
cells, e.g., tumor infiltration lymphocytes (TILs), such as T reg
cells, within the tumor.
In certain embodiments, the anti-GITR antibody used to determine
the level of GITR protein expression is an antibody comprising the
heavy and light chain variable region CDR sequences set forth in
SEQ ID NOs: 5-7 and 8-10, respectively, heavy and light chain
variable region sequences set forth in SEQ ID NOs: 11 and 12,
respectively, or full length heavy and light chain sequences of
antibody 6G10 (a fully human anti-GITR antibody) set forth in SEQ
ID NOs: 13 and 14, respectively, wherein the antibody or
antigen-binding fragment thereof specifically binds to human GITR.
In certain embodiments, the heavy chain lacks the C-terminal
lysine. In some embodiments, the antibody is a chimeric antibody
comprising the variable regions of antibody 6G10, but non-human
constant regions (e.g., mouse constant regions), e.g., a chimeric
6G10 antibody comprising the heavy and light chain sequences set
forth in SEQ ID NOs: 15 and 16, respectively. In certain
embodiments, the heavy chain of the chimeric antibody lacks the
C-terminal lysine. In a preferred embodiment, the detection of GITR
expression is performed by immunohistochemistry, as described
herein. In some embodiments, the level of GITR protein expression
is determined using a scoring system selected from the group
consisting of: the H score system, the Allred score system, an
AQUA.RTM. system, or an automated scoring system, as described
herein.
In certain embodiments, the anti-GITR immunotherapy comprises
administering to the cancer patient a therapeutically-effective
amount of an anti-GITR antibody disclosed in WO15187835, the
contents of which are herein incorporated by reference, e.g.,
antibodies having the heavy and light chain variable region CDRs,
heavy and light chain variable regions, or heavy and light chains
of antibodies 28F3, 19D3, 18E10, 3C3-1, 3C3-2, 2G6, 9G7-1, 9G7-2,
14E3, 19H8-1, 19H8-2, and/or 6G10, and variants thereof. The
sequences of the antibodies disclosed in PCT Patent Application No.
WO15187835 are provided in Table 2 (see SEQ ID NOs: 5-14 and
27-228). The patient may also be treated with any other anti-GITR
antibodies, e.g., TRX518 (Leap Therapeutics), MK-4166 (Merck),
LKZ-145 (Novartis), GWN-323 (Novartis Pharmaceuticals Corp.), Medi
1873 (MedImmune), INBRX-110 (Inhibrx), GITR-Fc protein (OncoMed)
and antibodies described in WO2006105021, WO2009009116,
WO2011028683, US2014/0072565, US20140072566, US20140065152,
WO2015031667, WO15184099, WO2015184099, or WO2016054638.
In certain embodiments, the status of a GITR-expressing tumor can
be monitored repeatedly after initial diagnosis (i.e., after making
a determination that the tumor is GITR-positive), such as one month
after initial diagnosis, two months after initial diagnosis, three
months after initial diagnosis, four months after initial
diagnosis, five months after initial diagnosis, six months after
initial diagnosis, one year after initial diagnosis, etc. In other
embodiments, the efficacy of anti-GITR immunotherapy against a
GITR-expressing tumor can be monitored repeatedly after initiating
anti-GITR immunotherapy, such as one month after initiation of
therapy, two months after initiation of therapy, three months after
initiation of therapy, four months after initiation of therapy,
five months after initiation of therapy, six months after
initiation of therapy, one year after initiation of therapy,
etc.
Diagnostic Kits
Provided herein are kits which include the diagnostic anti-GITR
antibodies described herein and instructions for use. Accordingly,
in some embodiments, the kit comprises an antibody comprising the
heavy and light chain variable region CDR sequences set forth in
SEQ ID NOs: 5-7 and 8-10, respectively, heavy and light chain
variable region sequences set forth in SEQ ID NOs: 11 and 12,
respectively, wherein the antibody or antigen-binding fragment
thereof specifically binds to human GITR, or full length heavy and
light chain sequences of antibody 6G10 set forth in SEQ ID NOs: 13
and 14, respectively. In certain embodiments, the heavy chain lacks
the C-terminal lysine. In some embodiments, the anti-GITR antibody
is a chimeric antibody comprising the variable regions of antibody
6G10, but non-human constant regions (e.g., mouse constant
regions), e.g., a chimeric antibody comprising the heavy and light
chain sequences set forth in SEQ ID NOs: 15 and 16, respectively.
In certain embodiments, the heavy chain of the chimeric antibody
lacks the C-terminal lysine. In certain embodiments, the antibody
comprises a detectable label, e.g., an antibody-label conjugate.
Such kits may comprise at least one additional reagent. For
example, in certain embodiments, the kits comprise buffers,
stabilizers, substrates, immunodetection reagent (e.g., secondary
antibodies for use in immunohistochemistry), and/or cofactors
required for the assay. In certain embodiments, the antibody and,
optionally the reagents, are suitably aliquoted. In certain
embodiments, the kit comprises a means for obtaining the biological
sample from a cancer patient. Such means can comprise, for example,
reagents that can be used to obtain fluid or tissue sample from the
cancer patient.
The present disclosure is further illustrated by the following
examples, which should not be construed as further limiting. The
contents of all figures and all references, Genbank sequences,
patents and published patent applications cited throughout this
application are expressly incorporated herein by reference. In
particular, the disclosure of WO15187835 is expressly incorporated
herein by reference.
EXAMPLES
Example 1: Generation of the 6G10 Antibody
The human anti-GITR antibody 6G10 was generated during the course
of immunizing HuMAb.RTM. transgenic mice ("HuMAb" is a Trade Mark
of Medarex, Inc., Princeton, N.J.) and KM mice (the KM Mouse.RTM.
strain contains the SC20 transchromosome as described in PCT
Publication WO 02/43478) and screening fusions for antibodies that
bound to GITR (see concurrently filed PCT Application No.
PCT/US15/33991). cDNA sequencing identified one heavy and one light
chain for the 6G10 antibody. The variable region amino acid
sequences and the isotype of the 6G10 antibody is set forth in
FIGS. 1-4 and Table 2.
Example 2: Binding Properties of the 6G10 Antibody
Binding of the 6G10 antibody to soluble GITR was determined by
Biacore. The 6G10 antibody was captured on human kappa coated chips
(.about.5KRUs; Southernbiotech), and recombinant human GITR
(rHGITR/Fc: R&D systems) was flowed across the chip at
concentrations of 500 nM, 250 nM, 125 nM, 62 nM, and 31 nM. The
capture concentration of the antibody/volume was 2-40 .mu.g/mL (5
.mu.L at 10 .mu.L/min). The antigen association time was 5 minutes
at 15 .mu.L/min, the antigen dissociation time was 6 minutes, and
regeneration was performed with 50 mM HCl/50 mM NaOH (12 .mu.L each
at 100 .mu.L/min). The 6G10 exhibited a K.sub.D of
1.87.times.10.sup.-9 M for binding to soluble human GITR, with a
k.sub.a of 3.83.times.10.sup.5 M.sup.-1 s.sup.-1 and a k.sub.d of
7.15.times.10.sup.-4 s.sup.-1.
Example 3: Immunohistochemistry of Cancer Tissue with a Chimeric
6G10 Antibody
Two immunohistochemistry (IHC) techniques were used to detect GITR
in tissue sections: one IHC method was conducted on formalin fixed
paraffin embedded (FFPE) tissue sections, and the other one was
conducted on frozen tissue sections.
Several anti-GITR antibodies have been tested in these IHC methods
and 6G10 was consistently better.
To detect GITR expression in human tissues, a fluoresceinated form
of the 6G10 antibody was prepared and designated 6G10-FITC. To
further develop a user and clinical friendly method, the 6G10
antibody was engineered to a mouse chimeric IgG2a format and
optimized for automated immunohistochemistry using the Leica BondRX
platform. The heavy and light chain sequences of the chimeric 6G10
antibody is set forth in FIGS. 5 and 6, respectively, and Table 2.
The heavy chain may or may not contain a C-terminal lysine.
Briefly, slides were deparaffinized and rehydrated following
routine histology procedures, and subjected to HIER (heat-induced
antigen retrieval) manually with pH6 target retrieval buffer (Dako)
for 10 min at 110.degree. C. using Decloaking Chamber (BioCare
Medical). Slides were then loaded onto the Leica BondRX automated
immunohistochemistry stainer. Tissue sections were incubated with
the chimeric 6G10 antibody at 0.5 .mu.g/ml for 60 minutes, followed
by detection with the Leica Bond Polymer Refine Detection System
(DS9800) using the manufacturer's instructions. Briefly, the tissue
sections were incubated with rabbit anti-mouse linker (Leica) for
30 minutes and Novolink Max polymer (Leica) for 30 minutes.
Finally, slides were reacted with a DAB substrate-chromogen
solution for 6 minutes. Slides were then counterstained with
Mayer's hematoxylin, dehydrated, cleared, and coverslipped with
Permount following routine histological procedure. Dako protein
block supplemented with 0.5% Hu-gamma-globulins (Sigma) was used to
block non-specific binding and PBS supplemented with 0.5% human
gamma globulins and 0.5% BSA was used as diluent for both primary
and secondary antibodies.
To profile tumor infiltrating lymphocytes (TILs), commercial
monoclonal antibodies to CD3 (T cell marker) and FoxP3 (regulatory
T cell marker) were used to stain adjacent sections. In-house
engineered mouse chimeric human anti-KLH IgG2a and commercial mouse
IgG1 were used as negative control antibodies. HEK-293T cells
stably expressing human GITR and hyperplasic human tonsil tissue
sections were used as positive controls. FFPE tissue samples from 5
tumor types were purchased from commercial tissue vendors, with
13.about.26 cases for each tumor type.
After immunostaining, the slides were examined under a light
microscope. For scoring of GITR and TILs, an automated and
quantitative analysis using Aperio.RTM. ScanScope and HALO.TM.
image analysis software was performed on the whole slide.
Representative images were taken either with Aperio ScanScope or
Olympus DP71 digital microscope camera.
Table 1 summarizes the distribution of GITR positive TILs in
cervical carcinoma (CC), melanoma (Mel), hepatocellular carcinoma
(HCC), colorectal carcinoma (CRC), and head and neck squamous cell
carcinoma (HNSCC). A manual score of 13 to 26 cases by estimation
of number of positive cells under 20.times. objective of
microscope. Minimum, <1 cells per 20.times. objective field;
Mild, 1.about.<10 cells per 20.times. objective field; Moderate,
10.about.<50 cells per 20.times. objective field; Marked,
50.about.<200 cells per 20.times. objective; Intense, >200
cells per 20.times. objective field. At least 1/3 of the whole
tumor section met the above criteria.
TABLE-US-00001 TABLE 1 Hepato- Head and neck Cervical Mela-
cellular Colorectal squamous cell Staining carcinoma noma carcinoma
carcinoma carcinoma intensity (n = 24) (n = 17) (n = 25) (n = 26)
(n = 13) Minimum 0% 6% 8% 12% 15% Mild 0% 24% 16% 15% 23% Moderate
58% 29% 36% 46% 38% Marked 33% 29% 40% 27% 24% Intense 8% 12% 0% 0%
0%
In hyperplastic tonsil, strong scattered positive cells were
distributed in the interfollicular region and germinal center, as
well as scattered epithelium-infiltrating lymphocytes. There was a
tendency for more positive cells to be distributed in focal
clusters of MNC beneath the epithelium. CD3+ TILs were present in
all cancer samples examined, although the quantity varied across
cancers and the distribution within the same tissue was
heterogeneous. CD8+ TILs were present in the vast majority of
tissues. In multiple cancers examined, GITR-positive staining was
present in a small fraction of TILs. In general, the abundance of
GITR+ cells was in proportion to that of TILs. GITR-positive TILs
were abundant in cervical carcinoma (CC), melanoma (Mel), and
hepatocellular carcinoma (HCC), and some staining was also observed
in colorectal carcinoma (CRC) and head and neck squamous cell
carcinoma (HNSCC), among which cervical squamous cell carcinoma
showed the most abundant staining.
GITR-positive staining was also observed in tumor cells in NSCLC
(non-small cell lung carcinoma), CC, HNSCC, and HCC (FIG. 7). In
NSCLC and CC, the specificity of the staining was confirmed by
complete displacement of staining upon incubation with GITR fusion
protein.
Immunohistochemistry was also performed on frozen tissue samples.
Cryostat sections at 5 .mu.m were fixed with acetone for 10 minutes
and immunostained using an indirect immunoperoxidase method at room
temperature. For staining with CD3 (T cell marker), CD8 (cytotoxic
T cell marker), and FoxP3 (regulatory T cell marker) monoclonal
antibodies, slides were further fixed with 10% NBF for 2 minutes.
For staining with 6G10-FITC and HuIgG2-FITC, slides were immersed
in 10% NBF for 1 minute. Peroxidase block supplied with the
EnVision+System and Dako protein block supplemented with 0.5% human
gamma globulins were used for endogenous peroxidase activity and
nonspecific binding blocks, respectively. Primary antibodies
(6G10-FITC at 5 .mu.g/mL, CD3 at 1:100, CD8 at 1:100 or FoxP3 at 5
or 10 .mu.g/mL) or an isotype control antibody (HuIgG2-FITC at 5
.mu.g/mL) were applied to sections and incubated for 1 hour. The
EnVison+system for either mouse or rabbit IgG was used as the
detection system. For 6G10-FITC and its isotype control
HuIgG2-FITC, rabbit anti-FITC antibody (5 .mu.g/mL) was used as
bridging antibody. Finally, slides were reacted with the DAB
substrate-chromogen solution for 6 minutes, and then counterstained
with Mayer's hematoxylin, dehydrated, cleared, and coverslipped
with Permount using routine histological procedures. For CD3, CD8,
and FoxP3 monoclonal antibodies, Dako protein block was used as the
diluent, whereas for 6G10-FITC and HuIgG2-FITC, PBS supplemented
with 0.5% human gamma globulins and 0.5% BSA was used as the
diluent for both primary and secondary antibodies. Frozen tissue
samples from 3 tumor types were purchased from commercial tissue
vendors.
Different amounts of GITR+ TILs were observed in the 3 tumor types.
FoxP3 was present in a small subset of TILs. Staining of a subset
of tumor cells with the anti-GITR antibody was also observed, and
the specificity of the staining was confirmed by complete
displacement of staining upon incubation with GITR fusion protein.
The presence of GITR mRNA in GITR tumor cells was also observed by
RNAscope on the frozen sections, which further confirms the IHC
results.
These results indicate that anti-GITR antibody 6G10 can efficiently
detect human GITR protein in fixed and frozen tissue samples.
TABLE-US-00002 TABLE 2 Summary table of sequences SEQ ID
DESCRIPTION SEQUENCE 1 human GITR isoform 1
MAQHGAMGAFRALCGLALLCALSLGQRPTGGPGCGPGRLLLG
TGTDARCCRVHTTRCCRDYPGEECCSEWDCMCVQPEFHCGDP
CCTTCRHHPCPPGQGVQSQGKFSFGFQCIDCASGTFSGGHEG
HCKPWTDCTQFGFLTVFPGNKTHNAVCVPGSPPAEPLGWLTV
VLLAVAACVLLLTSAQLGLHIWQLRSQCMWPRETQLLLEVPP
STEDARSCQFPEEERGERSAEEKGRLGDLWV 2 human GITR isoform 2
MAQHGAMGAFRALCGLALLCALSLGQRPTGGPGCGPGRLLLG
TGTDARCCRVHTTRCCRDYPGEECCSEWDCMCVQPEFHCGDP
CCTTCRHHPCPPGQGVQSQGKFSFGFQCIDCASGTFSGGHEG
HCKPWTDCCWRCRRRPKTPEAASSPRKSGASDRQRRRGGWET
CGCEPGRPPGPPTAASPSPGAPQAAGALRSALGRALLPWQQK
WVQEGGSDQRPGPCSSAAAAGPCRRERETQSWPPSSLAGPDG VGS 3 human GITR isoform
3 MAQHGAMGAFRALCGLALLCALSLGQRPTGGPGCGPGRLLLG
TGTDARCCRVHTTRCCRDYPGEECCSEWDCMCVQPEFHCGDP
CCTTCRHHPCPPGQGVQSQGKFSFGFQCIDCASGTFSGGHEG
HCKPWTDCTQFGFLTVFPGNKTHNAVCVPGSPPAEPLGWLTV
VLLAVAACVLLLTSAQLGLHIWQLRKTQLLLEVPPSTEDARS CQFPEEERGERSAEEKGRLGDLWV
4 extracellular portion QRPTGGPGCGPGRLLLGTGTDARCCRVHTTRCCRDYPGEECC
of mature SEWDCMCVQPEFHCGDPCCTTCRHHPCPPGQGVQSQGKFSFG human GITR
FQCIDCASGTFSGGHEGHCKPWTDCTQFGFLTVFPGNKTHNA VCVPGSPPAEP 5 6G10 VH
CDR1 TYGMH 6 6G10 VH CDR2 VTWYAGSNKFYADSVKG 7 6G10 VH CDR3
GGSMVRGLYYYGMDV 8 6G10 VL CDR1 RASQGISSALA 9 6G10 VL CDR2 DASSLES
10 6G10 VL CDR3 QQFNSYPYT 11 6G10 VH
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPG
KGLEWVAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMN
SLRAEDTAVYYCARGGSMVRGLYYYGMDVWGQGTTVTVSS 12 6G10 VL
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGK
APKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFA TYYCQQFNSYPYTFGQGTKLEIK
13 6G10 full length QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPG
heavy chain KGLEWVAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMN
SLRAEDTAVYYCARGGSMVRGLYYYGMDVWGQGTTVTVSSAS
TKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSG
ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVD
HKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKD
TLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKP
REEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIE
KTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS
DIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSLSLSPGK 14 6G10 full length
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGK light chain
APKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFA
TYYCQQFNSYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLK
SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS
KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFN RGEC 15 chimeric 6G10
antibody- QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPG full length
heavy chain KGLEWVAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMN The constant
region is SLRAEDTAVYYCARGGSMVRGLYYYGMDVWGQGTTVTVSSAK underlined
TTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSG
SLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNVAH
PASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIFPP
KIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVHTA
QTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLP
APIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTD
FMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLRVE
KKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK 16 chimeric 6G10 antibody-
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGK full length light chain
APKLLIYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFA The constant region is
TYYCQQFNSYPYTFGQGTKLEIKRADAAPTVSIFPPSSEQLT underlined
SGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDS
KDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFN RNEC 17 6G10 (VH)
nucleotide CAGGTGCAGCTGGTGGAGTCTGGGGGAGACGTGGT sequence
CCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAG
CGTCTGGATTCACCTTCAGTACCTATGGCATGCAC
TGGGTCCGCCAGGCTCCAGGCAAGGGGCTGGAGTG
GGTGGCAGTTACATGGTATGCTGGAAGTAATAAAT
TTTATGCAGACTCCGTGAAGGGCCGATTCACCATC
TCCAGAGACAATTCCAAGAACACGCTGTATCTGCA
AATGAACAGCCTGAGAGCCGAGGACACGGCTGTGT
ATTACTGTGCGAGAGGAGGTAGTATGGTTCGGGGA
CTTTATTATTACGGTATGGACGTCTGGGGCCAAGG GACCACGGTCACCGTCTCCTCA 18 6G10
(VL) nucleotide GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTC sequence
TGCATCTGTAGGAGACAGAGTCACCATCACTTGCC
GGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGG
TATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCT
GATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCC
CATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAT
TTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGA
TTTTGCAACTTATTACTGTCAACAGTTTAATAGTT
ACCCGTACACTTTTGGCCAGGGGACCAAGCTGGAG ATCAAA 19 6G10 (full length
wild- CAGGTGCAGCTGGTGGAGTCTGGGGGAGACGTGGTCCAGCCT type heavy chain)
GGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACC nucleotide sequence
TTCAGTACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGC The sequence encoding
AAGGGGCTGGAGTGGGTGGCAGTTACATGGTATGCTGGAAGT the constant region
AATAAATTTTATGCAGACTCCGTGAAGGGCCGATTCACCATC is underlined
TCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGA
GGAGGTAGTATGGTTCGGGGACTTTATTATTACGGTATGGAC
GTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCTCC
ACCAAGGGCCCATCGGTCTTCCCCCTGGCGCCCTGCTCCAGG
AGCACCTCCGAGAGCACAGCGGCCCTGGGCTGCCTGGTCAAG
GACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGC
GCTCTGACCAGCGGCGTGCACACCTTCCCAGCTGTCCTACAG
TCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCC
TCCAGCAACTTCGGCACCCAGACCTACACCTGCAACGTAGAT
CACAAGCCCAGCAACACCAAGGTGGACAAGACAGTTGAGCGC
AAATGTTGTGTCGAGTGCCCACCGTGCCCAGCACCACCTGTG
GCAGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC
ACCCTCATGATCTCCCGGACCCCTGAGGTCACGTGCGTGGTG
GTGGACGTGAGCCACGAAGACCCCGAGGTCCAGTTCAACTGG
TACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCA
CGGGAGGAGCAGTTCAACAGCACGTTCCGTGTGGTCAGCGTC
CTCACCGTTGTGCACCAGGACTGGCTGAACGGCAAGGAGTAC
AAGTGCAAGGTCTCCAACAAAGGCCTCCCAGCCCCCATCGAG
AAAACCATCTCCAAAACCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAAC
CAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTACCCCAGC
GACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAAC
AACTACAAGACCACACCTCCCATGCTGGACTCCGACGGCTCC
TTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGG
CAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCT
CTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCG GGTAAA 20 6G10 (full
length wild- GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCT type light
chain) GTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC nucleotide
sequence ATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAA The sequence
encoding GCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGT the constant
region GGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAT is underlined
TTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCA
ACTTATTACTGTCAACAGTTTAATAGTTACCCGTACACTTTT
GGCCAGGGGACCAAGCTGGAGATCAAACGAACTGTGGCTGCA
CCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAA
TCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTAT
CCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTC
CAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGC
AAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGC
AAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTC
ACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAAC AGGGGAGAGTGT 21 chimeric
6G10 heavy CAGGTGCAGCTGGTGGAGTCTGGGGGAGACGTGGTCCAGCCT chain
GGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACC The sequence encoding
TTCAGTACCTATGGCATGCACTGGGTCCGCCAGGCTCCAGGC the constant region
AAGGGGCTGGAGTGGGTGGCAGTTACATGGTATGCTGGAAGT is underlined
AATAAATTTTATGCAGACTCCGTGAAGGGCCGATTCACCATC
TCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAAC
AGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGA
GGAGGTAGTATGGTTCGGGGACTTTATTATTACGGTATGGAC
GTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCCAAA
ACAACAGCCCCATCGGTCTATCCGCTAGCCCCTGTGTGTGGA
GATACAACTGGCTCCTCGGTGACTCTAGGATGCCTGGTCAAG
GGTTATTTCCCTGAGCCAGTGACCTTGACCTGGAACTCTGGA
TCCCTGTCCAGTGGTGTGCACACCTTCCCAGCTGTCCTGCAG
TCTGACCTCTACACCCTCAGCAGCTCAGTGACTGTAACCTCG
AGCACCTGGCCCAGCCAGTCCATCACCTGCAATGTGGCCCAC
CCGGCAAGCAGCACCAAGGTGGACAAGAAAATTGAGCCCAGA
GGGCCCACAATCAAGCCCTGTCCTCCATGCAAATGCCCAGCA
CCTAACCTCTTGGGTGGACCATCCGTCTTCATCTTCCCTCCA
AAGATCAAGGATGTACTCATGATCTCCCTGAGCCCCATAGTC
ACATGTGTGGTGGTGGATGTGAGCGAGGATGACCCAGATGTC
CAGATCAGCTGGTTTGTGAACAACGTGGAAGTACACACAGCT
CAGACACAAACCCATAGAGAGGATTACAACAGTACTCTCCGG
GTGGTCAGTGCCCTCCCCATCCAGCACCAGGACTGGATGAGT
GGCAAGGAGTTCAAATGCAAGGTCAACAACAAAGACCTCCCA
GCGCCCATCGAGAGAACCATCTCAAAACCCAAAGGGTCAGTA
AGAGCTCCACAGGTATATGTCTTGCCTCCACCAGAAGAAGAG
ATGACTAAGAAACAGGTCACTCTGACCTGCATGGTCACAGAC
TTCATGCCTGAAGACATTTACGTGGAGTGGACCAACAACGGG
AAAACAGAGCTAAACTACAAGAACACTGAACCAGTCCTGGAC
TCTGATGGTTCTTACTTCATGTACAGCAAGCTGAGAGTGGAA
AAGAAGAACTGGGTGGAAAGAAATAGCTACTCCTGTTCAGTG
GTCCACGAGGGTCTGCACAATCACCACACGACTAAGAGCTTC TCCCGGACTCCGGGTAAATGA 22
chimeric 6G10 light GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCT
chain GTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGC The sequence
encoding ATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAA the constant
region GCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGT is underlined
GGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAT
TTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCA
ACTTATTACTGTCAACAGTTTAATAGTTACCCGTACACTTTT
GGCCAGGGGACCAAGCTGGAGATCAAACGGGCTGATGCTGCA
CCAACTGTATCCATCTTCCCACCATCCAGTGAGCAATTGACA
TCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTTCTAC
CCCAAAGACATCAATGTCAAGTGGAAGATTGATGGCAGTGAA
CGACAAAATGGCGTCCTGAACAGTTGGACCGATCAGGACAGC
AAAGACAGCACCTACAGCATGAGCAGCACCCTCACGTTGACC
AAGGACGAGTATGAACGACATAACAGCTATACCTGTGAGGCC
ACTCACAAGACATCAACTTCACCCATTGTCAAGAGCTTCAAC AGGAATGAGTGTTAG 23 human
IgG2 heavy chain ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWN
constant region SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCN
VDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKP
KDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKT
KPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAP
IEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKS
RWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 24 human kappa light chain
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK constant region
VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKV YACEVTHQGLSSPVTKSFNRGEC
25 mouse IgG2a heavy chain
AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWN constant region
SGSLSSGVHTFPAVLQSDLYTLSSSVTVTSSTWPSQSITCNV
AHPASSTKVDKKIEPRGPTIKPCPPCKCPAPNLLGGPSVFIF
PPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVH
TAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKD
LPAPIERTISKPKGSVRAPQVYVLPPPEEEMTKKQVTLTCMV
TDFMPEDIYVEWTNNGKTELNYKNTEPVLDSDGSYFMYSKLR
VEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPGK 26 mouse kappa light chain
RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWK constant region
IDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNS YTCEATHKTSTSPIVKSFNRNEC
27 28F3 (VH) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGDYYYGMDVWGQGTTVTVSS 28 28F3 (VL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY PYTFGQGTKLEIK
29 28F3 (full length wild-
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW type heavy chain)
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY The constant region
is YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS underlined
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPGK 30 28F3 (full length wild-
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLL type light chain)
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 31 28F3.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 32 28F3.IgG1.1
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW (VH + IgG1.1)
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 33 28F3.IgG1 (VL + CL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 34 28F3 VH CDR1 SYGMH 35 28F3 VH CDR2
VIWYEGSNKYYADSVKG 36 28F3 VH CDR3 GGSMVRGDYYYGMDV 37 28F3 VL CDR1
RASQGISSALA 38 28F3 VL CDR2 DASSLES 39 28F3 VL CDR3 QQFNSYPYT 40
28F3 (VH + G2(C219S))
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW or
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY 28F3-IgG2-C219S
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 41 28F3 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW 28F3-IgG2-IgG1
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 42 28F3 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW 28F3-IgG2-IgG1.1
VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 43 28F3
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW (VH + G2(C219S).g1)
or VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
28F3-IgG2-C219S-IgG1
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 44 28F3
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW (VH +
G2(C219S).g1.1) VAVIWYEGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY or
YCARGGSMVRGDYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
28F3-IgG2-C219S-IgG1.1
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 45 28F3 (VH)
MRAWIFFLLCLAGRALAQVQLVESGGGVVQPGRSLRLSCAASGFTFS (SEQ ID NO: 13)
SYGMHWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRFTISRDNSKN with signal peptide
TLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVSS The signal peptide
is underlined 46 28F3 (VL)
MRAWIFFLLCLAGRALAAIQLTQSPSSLSASVGDRVTITCRASQGIS (SEQ ID NO: 14)
SALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISS with signal peptide
LQPEDFATYYCQQFNSYPYTFGQGTKLEIK The signal peptide is underlined 47
28F3.IgG1 (VH + IgG1)
MRAWIFFLLCLAGRALAQVQLVESGGGVVQPGRSLRLSCAASGFTFS with signal peptide
SYGMHWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRFTISRDNSKN The signal peptide
TLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVSS and constant region
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT are underlined
SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV
DKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPE
VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 48 28F3.IgG1.1
MRAWIFFLLCLAGRALAQVQLVESGGGVVQPGRSLRLSCAASGFTFS (VH + IgG1.1)
SYGMHWVRQAPGKGLEWVAVIWYEGSNKYYADSVKGRFTISRDNSKN with signal peptide
TLYLQMNSLRAEDTAVYYCARGGSMVRGDYYYGMDVWGQGTTVTVSS The signal
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT peptide and
constant SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV region are
underlined DKRVEPKSCDKTHTCPPCPAPEAEGAPSVFLFPPKPKDTLMISRTPE
VTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV
LTVLHQDWLNGKEYKCKVSNKALPSSIEKTISKAKGQPREPQVYTLP
PSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD
SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 49 28F3.IgG1 (VL +
CL) with signal MRAWIFFLLCLAGRALAAIQLTQSPSSLSASVGDRVTITCRASQGIS
peptide SALAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTDFTLTISS The signal
peptide and constant
LQPEDFATYYCQQFNSYPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQ region are
underlined LKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS
TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 50 19D3 (VH)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSS 51 19D3 (VL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY PYTFGQGTKLEIK 52
19D3 (full length wild-type
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW heavy chain)
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY The constant region
is YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST underlined
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVD
GVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPGK 53 19D3 (full length wild-type
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL light chain)
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 54 19D3.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSLSLSPG 55 19D3.IgG1.1 (VH + IgG1.1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC
PAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSLSLSPG 56 19D3.IgG1 (VL + CL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 57 19D3 VH CDR1 SYGFH 58 19D3 VH CDR2
VIWYAGSNKFYADSVKG 59 19D3 VH CDR3 GGQLDYYYYYVMDV 60 19D3 VL CDR1
RASQGISSWLA 61 19D3 VL CDR2 AASSLQS 62 19D3 VL CDR3 QQYNSYPYT 63
19D3 (VH + G2(C219S)) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-C219S
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVD
GVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 64 19D3 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-IgG1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 65 19D3 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-IgG1.1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 66 19D3 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
19D3-IgG2-C219S-IgG1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 67 19D3 (VH + G2(C219S).g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
19D3-IgG2-C219S-IgG1.1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 68 19D3 (VH + G2(C219S)) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-C219S
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVD
GVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 69 19D3 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-IgG1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 70 19D3 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW 19D3-IgG2-IgG1.1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 71 19D3 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
19D3-IgG2-C219S-IgG1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 72 19D3 (VH + G2(C219S).g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGFHWVRQAPGKGLEW
19D3-IgG2-C219S-IgG1.1
VAVIWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGQLDYYYYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 73 18E10 (VH)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSS 74 18E10 (VL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY PYTFGQGTKLEIK 75
18E10 (full length wild-type
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW heavy chain)
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY The constant region
is YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST underlined
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVD
GVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPGK 76 18E10 (full length wild-type
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL light chain)
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 77 18E10.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC
PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSLSLSPG 78 18E10.IgG1.1 (VH + IgG1.1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPSSKST
SGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC
PAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFN
WYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKV
SNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK
GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRW
QQGNVFSCSVMHEALHNHYTQKSLSLSPG 79 18E10.IgG1 (VL + CL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 80 18E10 VH CDR1 SYGMH 81 18E10 VH CDR2
VIWYAGSNKYYADSVKG 82 18E10 VH CDR3 GGRIAVAFYYSMDV 83 18E10 VL CDR1
RASQGISSWLA 84 18E10 VL CDR2 AASSLQS 85 18E10 VL CDR3 QQYNSYPYT 86
18E10 (VH + G2(C219S)) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW 18E10-IgG2-C219S
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVD
GVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKG
LPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 87 18E10 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW 18E10-IgG2-IgG1
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 88 18E10 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW 18E10-IgG2-IgG1.1
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 89 18E10 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW
18E10-IgG2-C219S-IgG1
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 90 18E10 (VH + G2(C219S).g1.1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQAPGKGLEW or
VAVIWYAGSNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
18E10-IgG2-C219S-IgG1.1
YCARGGRIAVAFYYSMDVWGQGTTVTVSSASTKGPSVFPLAPCSRST
SESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL
SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAP
PVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVD
GVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYP
SDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGN
VFSCSVMHEALHNHYTQKSLSLSPG 91 3C3 (VH)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSS 92 3C3 (VL1)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY PYTFGQGTKLEIK 93
3C3 (VL2) EIVLTQSPATLSLSPGERATLSCRASQGVSSYLAWYQQKPGQAPRLL
IYDASNRATGIPARFSGSGPGTDFTLTISSLEPEDFAVYYCQQRSNW HTFGQGTKLEIK 94 3C3
(full length wild-type
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW heavy chain)
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY The constant region
is CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA underlined
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL
GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPGK 95 3C3 L1 (full length wild-type
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL light chain 1)
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC
96 3C3 L2 (full length wild-type
EIVLTQSPATLSLSPGERATLSCRASQGVSSYLAWYQQKPGQAPRLL light chain 2)
IYDASNRATGIPARFSGSGPGTDFTLTISSLEPEDFAVYYCQQRSNW The constant region
is HTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR underlined
EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 97 3C3.IgG1 (VH + IgG1)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL
GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPG 98 3C3.IgG1.1 (VH + IgG1.1)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAE
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
SSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPG 99 3C3.IgG1 (VL1 + CL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 100 3C3IgG1.2 (VL2 + CL)
EIVLTQSPATLSLSPGERATLSCRASQGVSSYLAWYQQKPGQAPRLL
IYDASNRATGIPARFSGSGPGTDFTLTISSLEPEDFAVYYCQQRSNW
HTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR
EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH
KVYACEVTHQGLSSPVTKSFNRGEC 101 3C3 VH CDR1 GYYWT 102 3C3 VH CDR2
KINHSGNTNYNPSLKS 103 3C3 VH CDR3 LGAFDAFDI 104 3C3 VL1 CDR1
RASQGISSWLA 105 3C3 VL1 CDR2 AASSLQS 106 3C3 VL1 CDR3 QQYNSYPYT 107
3C3 VL2 CDR1 RASQGVSSYLA 108 3C3 VL2 CDR2 DASNRAT 109 3C3 VL2 CDR3
QQRSNWHT 110 3C3 (VH + G2) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW 3C3-IgG2
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHN
AKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIE
KTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
111 3C3 (VH + G2(C219S)) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW 3C3-IgG2-C219S
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHN
AKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIE
KTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
112 3C3 (VH + G2.g1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW 3C3-IgG2-IgG1
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPELLGGP
SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV
EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPG 113 3C3 (VH + G2.g1.1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW 3C3-IgG2-IgG1.1
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN
AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
114 3C3 (VH + G2(C219S).g1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW 3C3-IgG2-C219S-IgG1
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPELLGGP
SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV
EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPG 115 3C3 (VH + G2(C219S).g1.1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWTWIRQPPGKGLEW
3C3-IgG2-C219S-IgG1.1
IGKINHSGNTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYY
CARLGAFDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN
AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
116 2G6 (VH) QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSS 117 2G6 (VL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY PYTFGQGTKLEIK 118
2G6 (full length wild-type
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW heavy chain)
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY The constant region
is YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS underlined
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPGK 119 2G6 (full length wild-type
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL light chain)
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 120 2G6.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 121 2G6.IgG1.1 (VH + IgG1.1)
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 122 2G6.IgG1 (VL + CL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 123 2G6 VH CDR1 DYGMH 124 2G6 VH CDR2
VIWYDGSNKFYVDSVKG 125 2G6 VH CDR3 GGRLATGHFYYVMDV 126 2G6 VL CDR1
RASQGISSWLA 127 2G6 VL CDR2 AASSLQS 128 2G6 VL CDR3 QQYNSYPYT 129
2G6 (VH + G2) or QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW
2G6-IgG2 VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 130 2G6 (VH + G2(C219S)) or
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW 2G6-IgG2-C219S
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 131 2G6 (VH + G2.g1) or
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW 2G6-IgG2-IgG1
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 132 2G6 (VH + G2.g1.1) or
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW 2G6-IgG2-IgG1.1
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 133 2G6 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW 2G6-IgG2-C219S-IgG1
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 134 2G6 (VH + G2(C219S).g1.1) or
QVQLVESGGGVVQPGGSLRLSCAASGFILSDYGMHWVRQAPGKGLEW
2G6-IgG2-C219S-IgG1.1
VTVIWYDGSNKFYVDSVKGRFTISRDNSKNTLYLQMNSLRVEDTAVY
YCARGGRLATGHFYYVMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 135 8A6 (VH)
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSS 136 8A6 (VL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY PYTFGQGTKLEIK 137
8A6 (full length wild-type
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW heavy chain)
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY The constant region
is YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS underlined
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPGK 138 8A6 (full length wild-type
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL light chain)
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY The constant region
is PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 139 8A6.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 140 8A6.IgG1.1 (VH + IgG1.1)
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 141 8A6.IgG1 (VL + CL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 142 8A6 VH CDR1 SYGMQ 143 8A6 VH CDR2
VIWYEGSNKYYADSVKG 144 8A6 VH CDR3 GGLMVRGLFYYGMDV 145 8A6 VL CDR1
RASQGISSALA 146 8A6 VL CDR2 DASSLES 147 8A6 VL CDR3 QQFNSYPYT 148
8A6 (VH + G2(C219S)) or
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW 8A6-IgG2-C219S
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 149 8A6 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW 8A6-IgG2-IgG1
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 150 8A6 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW 8A6-IgG2-IgG1.1
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 151 8A6 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW 8A6-IgG2-C219S-IgG1
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 152 8A6 (VH + G2(C219S).g1.1) or
QVQLVESGGGVVQPGRSLRLSCTASGFTFSSYGMQWVRQAPGKGLEW
8A6-IgG2-C219S-IgG1.1
VAVIWYEGSNKYYADSVKGRFTISRENSKNTLYLQMNSLRAEDTAVY
YCARGGLMVRGLFYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 153 9G7 (VH)
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSS 154 9G7 (VL1)
EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRL
LIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS SPWTFGQGTKVEIK 155
9G7 (VL2) EIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRL
LIYGASSRATGIPERFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS SPITFGQGTRLEIK 156
9G7 (full length wild-type
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW heavy chain)
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA The constant region
is VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR underlined
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCP
APEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNW
YVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQ
EGNVFSCSVMHEALHNHYTQKSLSLSLGK 157 9G7 L2 (full length wild-type
EIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRL light chain 2)
LIYGASSRATGIPERFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS The constant region
is SPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY underlined
PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNRGEC 158 9G7.IgG1 (VH + IgG1)
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCP
PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS
RWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 159 9G7.IgG1.1 (VH + IgG1.1)
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPSSK
STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCP
PCPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVK
FNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKC
KVSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCL
VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKS
RWQQGNVFSCSVMHEALHNHYTQKSLSLSPG 160 9G7.IgG1 (VL1 + CL)
EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRL
LIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS
SPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY
PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNRGEC 161 9G7.IgG1.2 (VL2 + CL)
EIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRL
LIYGASSRATGIPERFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS
SPITFGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY
PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNRGEC 162 9G7 VH CDR1 TVWMS 163 9G7 VH CDR2
RIKSKTDGGTTDYAAPVKG 164 9G7 VH CDR3 GQLIPYSYYYGMDV 165 9G7 VL1 CDR1
RASQSVSSSYLA 166 9G7 VL1 CDR2 GASSRAT 167 9G7 VL1 CDR3 QQYGSSPWT
168 9G7 VL2 CDR1 RASQSVTSSYLA 169 9G7 VL2 CDR2 GASSRAT 170 9G7 VL2
CDR3 QQYGSSPIT 171 9G7 (VH + G2) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW 9G7-IgG2
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCP
APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWY
VDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSN
KGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 172 9G7 (VH + G2(C219S)) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW 9G7-IgG2-C219S
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCP
APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWY
VDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSN
KGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 173 9G7 (VH + G2.g1) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW 9G7-IgG2-IgG1
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ
QGNVFSCSVMHEALHNHYTQKSLSLSPG 174 9G7 (VH + G2.g1.1) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW 9G7-IgG2-IgG1.1
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCP
APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 175 9G7 (VH + G2(C219S).g1) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW 9G7-IgG2-C219S-IgG1
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCP
APELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW
YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS
NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKG
FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ
QGNVFSCSVMHEALHNHYTQKSLSLSPG 176 9G7 (VH + G2(C219S).g1.1) or
EVQLVESGGGLVKPGGSLRLSCAASGFTFSTVWMSWVRQAPGKGLEW
9G7-IgG2-C219S-IgG1.1
VGRIKSKTDGGTTDYAAPVKGRFTISRDDSKNTLYLQMNSLHTEDTA
VYYCTTGQLIPYSYYYGMDVWGQGTSVTVSSASTKGPSVFPLAPCSR
STSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY
SLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCP
APPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 177 9G7 L1 (full length wild-type
EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRL light chain 1)
LIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGS The constant region
is SPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY underlined
PREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE
KHKVYACEVTHQGLSSPVTKSFNRGEC 178 14E3 (VH)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSS 179 14E3 (VL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY PPTFGQGTKVEIK 180
14E3 (full length wild-type
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW heavy chain)
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY The constant region
is CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA underlined
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL
GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPGK 181 14E3 (full length wild-type
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL light chain)
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY The constant region
is PPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 182 14E3.IgG1 (VH + IgG1)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELL
GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
APIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPG 183 14E3.IgG1.1 (VH + IgG1.1)
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAE
GAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV
EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP
SSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD
IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF
SCSVMHEALHNHYTQKSLSLSPG 184 14E3.IgG1 (VL + CL)
DIQMTQSPSSLSASVGDRVTITCRASQGISSWLAWYQQKPEKAPKSL
IYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSY
PPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 185 14E3 VH CDR1 GYYWS 186 14E3 VH CDR2
EINHSGNTYYNPSLKS 187 14E3 VH CDR3 FGSNDAFDI 188 14E3 VL CDR1
RASQGISSWLA 189 14E3 VL CDR2 AASSLQS 190 14E3 VL CDR3 QQYNSYPPT 191
14E3 (VH + G2) or QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
14E3-IgG2 IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHN
AKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIE
KTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
192 14E3 (VH + G2(C219S)) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW 14E3-IgG2-C219S
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHN
AKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIE
KTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
193 14E3 (VH + G2.g1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW 14E3-IgG2-IgG1
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPELLGGP
SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV
EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPG 194 14E3 (VH + G2.g1.1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW 14E3-IgG2-IgG1.1
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN
AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
195 14E3 (VH + G2(C219S).g1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
14E3-IgG2-C219S-IgG1
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPELLGGP
SVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVH
NAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPI
EKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAV
EWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCS
VMHEALHNHYTQKSLSLSPG 196 14E3 (VH + G2(C219S).g1.1) or
QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEW
14E3-IgG2-C219S-IgG1.1
IGEINHSGNTYYNPSLKSRVTISVDTSKNQLSLKLSSVTAADTAVYY
CARFGSNDAFDIWGQGTMVTVSSASTKGPSVFPLAPCSRSTSESTAA
LGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV
PSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPAPPVAGPS
VFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN
AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPSSIE
KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE
WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPG
197 19H8 (VH) QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSS 198 19H8 (VL1)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY PQTFGQGTKVEIK 199
19H8 (VL2) EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL
IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNW PLTFGGGTKVEIK 200
19H8 (full length wild-type
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW heavy chain)
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY The constant region
is YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS underlined
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPGK 201 19H8 L1 (full length wild-type
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL light chain 1)
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY The constant region
is PQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP underlined
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 202 19H8 L2 (full length wild-type
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL light chain 2)
IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNW
PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 203 19H8.IgG1 (VH + IgG1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 204 19H8.IgG1.1 (VH + IgG1.1)
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 205 19H8.IgG1 (VL1 + CL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKFL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY
PQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 206 19H8.IgG1.2 (VL2 + CL)
EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLL
IYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNW
PLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 207 19H8 VH CDR1 NYGMH 208 19H8 VH CDR2
VIWYGGSNKFYADSVKG 209 19H8 VH CDR3 GGAMVRGVYYYGMDV 210 19H8 VL1
CDR1 RASQGISSALA 211 19H8 VL1 CDR2 DASSLES 212 19H8 VL1 CDR3
QQFNSYPQT 213 19H8 VL2 CDR1 RASQSVSSYLA 214 19H8 VL2 CDR2 DASNRAT
215 19H8 VL2 CDR3 QQRSNWPLT 216 19H8 (VH + G2(C219S)) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW 19H8-IgG2-C219S
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 217 19H8 (VH + G2.g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW 19H8-IgG2-IgG1
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 218 19H8 (VH + G2.g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW 19H8-IgG2-IgG1.1
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 219 19H8 (VH + G2(C219S).g1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW
19H8-IgG2-C219S-IgG1
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 220 19H8 (VH + G2(C219S).g1.1) or
QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYGMHWVRQAPGKGLEW
19H8-IgG2-C219S-IgG1.1
MAVIWYGGSNKFYADSVKGRFTISRDNSKNSLSLQMNSLRAEDTAVY
YCARGGAMVRGVYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 221 6G10.IgG1 (VH + IgG1)
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 222 6G10.IgG1.1 (VH + IgG1.1)
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKS
TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP
CPAPEAEGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKF
NWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCK
VSNKALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV
KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSR
WQQGNVFSCSVMHEALHNHYTQKSLSLSPG 223 6G10.IgG1 (VL + CL)
AIQLTQSPSSLSASVGDRVTITCRASQGISSALAWYQQKPGKAPKLL
IYDASSLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSY
PYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYP
REAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK
HKVYACEVTHQGLSSPVTKSFNRGEC 224 6G10 (VH + G2(C219S)) or
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW 6G10-IgG2-C219S
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYV
DGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNK
GLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 225 6G10 (VH + G2.g1) or 6G10-
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW IgG2-IgG1
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 226 6G10 (VH + G2.g1.1) or 6G10-
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW IgG2-IgG1.1
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG 227 6G10 (VH + G2(C219S).g1) or
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW
6G10-IgG2-C219S-IgG1
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWY
VDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN
KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGF
YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ
GNVFSCSVMHEALHNHYTQKSLSLSPG 228 6G10 (VH + G2(C219S).g1.1) or
QVQLVESGGDVVQPGRSLRLSCAASGFTFSTYGMHWVRQAPGKGLEW
6G10-IgG2-C219S-IgG1.1
VAVTWYAGSNKFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVY
YCARGGSMVRGLYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPCSRS
TSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS
LSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKSCVECPPCPA
PPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYV
DGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNK
ALPSSIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY
PSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQG
NVFSCSVMHEALHNHYTQKSLSLSPG
Table 2 provides the sequences of the mature variable regions and
heavy and light chains and where indicated, sequences with signal
peptides.
EQUIVALENTS
Those skilled in the art will recognize, or be able to ascertain
using no more than routine experimentation, many equivalents of the
specific embodiments disclosed herein. Such equivalents are
intended to be encompassed by the following claims
SEQUENCE LISTINGS
1
2281241PRTHomo sapiensmisc_feature(1)..(241)human GITR isoform 1
1Met Ala Gln His Gly Ala Met Gly Ala Phe Arg Ala Leu Cys Gly Leu1 5
10 15Ala Leu Leu Cys Ala Leu Ser Leu Gly Gln Arg Pro Thr Gly Gly
Pro 20 25 30Gly Cys Gly Pro Gly Arg Leu Leu Leu Gly Thr Gly Thr Asp
Ala Arg 35 40 45Cys Cys Arg Val His Thr Thr Arg Cys Cys Arg Asp Tyr
Pro Gly Glu 50 55 60Glu Cys Cys Ser Glu Trp Asp Cys Met Cys Val Gln
Pro Glu Phe His65 70 75 80Cys Gly Asp Pro Cys Cys Thr Thr Cys Arg
His His Pro Cys Pro Pro 85 90 95Gly Gln Gly Val Gln Ser Gln Gly Lys
Phe Ser Phe Gly Phe Gln Cys 100 105 110Ile Asp Cys Ala Ser Gly Thr
Phe Ser Gly Gly His Glu Gly His Cys 115 120 125Lys Pro Trp Thr Asp
Cys Thr Gln Phe Gly Phe Leu Thr Val Phe Pro 130 135 140Gly Asn Lys
Thr His Asn Ala Val Cys Val Pro Gly Ser Pro Pro Ala145 150 155
160Glu Pro Leu Gly Trp Leu Thr Val Val Leu Leu Ala Val Ala Ala Cys
165 170 175Val Leu Leu Leu Thr Ser Ala Gln Leu Gly Leu His Ile Trp
Gln Leu 180 185 190Arg Ser Gln Cys Met Trp Pro Arg Glu Thr Gln Leu
Leu Leu Glu Val 195 200 205Pro Pro Ser Thr Glu Asp Ala Arg Ser Cys
Gln Phe Pro Glu Glu Glu 210 215 220Arg Gly Glu Arg Ser Ala Glu Glu
Lys Gly Arg Leu Gly Asp Leu Trp225 230 235 240Val2255PRTHomo
sapiensmisc_feature(1)..(255)human GITR isoform 2 2Met Ala Gln His
Gly Ala Met Gly Ala Phe Arg Ala Leu Cys Gly Leu1 5 10 15Ala Leu Leu
Cys Ala Leu Ser Leu Gly Gln Arg Pro Thr Gly Gly Pro 20 25 30Gly Cys
Gly Pro Gly Arg Leu Leu Leu Gly Thr Gly Thr Asp Ala Arg 35 40 45Cys
Cys Arg Val His Thr Thr Arg Cys Cys Arg Asp Tyr Pro Gly Glu 50 55
60Glu Cys Cys Ser Glu Trp Asp Cys Met Cys Val Gln Pro Glu Phe His65
70 75 80Cys Gly Asp Pro Cys Cys Thr Thr Cys Arg His His Pro Cys Pro
Pro 85 90 95Gly Gln Gly Val Gln Ser Gln Gly Lys Phe Ser Phe Gly Phe
Gln Cys 100 105 110Ile Asp Cys Ala Ser Gly Thr Phe Ser Gly Gly His
Glu Gly His Cys 115 120 125Lys Pro Trp Thr Asp Cys Cys Trp Arg Cys
Arg Arg Arg Pro Lys Thr 130 135 140Pro Glu Ala Ala Ser Ser Pro Arg
Lys Ser Gly Ala Ser Asp Arg Gln145 150 155 160Arg Arg Arg Gly Gly
Trp Glu Thr Cys Gly Cys Glu Pro Gly Arg Pro 165 170 175Pro Gly Pro
Pro Thr Ala Ala Ser Pro Ser Pro Gly Ala Pro Gln Ala 180 185 190Ala
Gly Ala Leu Arg Ser Ala Leu Gly Arg Ala Leu Leu Pro Trp Gln 195 200
205Gln Lys Trp Val Gln Glu Gly Gly Ser Asp Gln Arg Pro Gly Pro Cys
210 215 220Ser Ser Ala Ala Ala Ala Gly Pro Cys Arg Arg Glu Arg Glu
Thr Gln225 230 235 240Ser Trp Pro Pro Ser Ser Leu Ala Gly Pro Asp
Gly Val Gly Ser 245 250 2553234PRTHomo
sapiensmisc_feature(1)..(234)human GITR isoform 3 3Met Ala Gln His
Gly Ala Met Gly Ala Phe Arg Ala Leu Cys Gly Leu1 5 10 15Ala Leu Leu
Cys Ala Leu Ser Leu Gly Gln Arg Pro Thr Gly Gly Pro 20 25 30Gly Cys
Gly Pro Gly Arg Leu Leu Leu Gly Thr Gly Thr Asp Ala Arg 35 40 45Cys
Cys Arg Val His Thr Thr Arg Cys Cys Arg Asp Tyr Pro Gly Glu 50 55
60Glu Cys Cys Ser Glu Trp Asp Cys Met Cys Val Gln Pro Glu Phe His65
70 75 80Cys Gly Asp Pro Cys Cys Thr Thr Cys Arg His His Pro Cys Pro
Pro 85 90 95Gly Gln Gly Val Gln Ser Gln Gly Lys Phe Ser Phe Gly Phe
Gln Cys 100 105 110Ile Asp Cys Ala Ser Gly Thr Phe Ser Gly Gly His
Glu Gly His Cys 115 120 125Lys Pro Trp Thr Asp Cys Thr Gln Phe Gly
Phe Leu Thr Val Phe Pro 130 135 140Gly Asn Lys Thr His Asn Ala Val
Cys Val Pro Gly Ser Pro Pro Ala145 150 155 160Glu Pro Leu Gly Trp
Leu Thr Val Val Leu Leu Ala Val Ala Ala Cys 165 170 175Val Leu Leu
Leu Thr Ser Ala Gln Leu Gly Leu His Ile Trp Gln Leu 180 185 190Arg
Lys Thr Gln Leu Leu Leu Glu Val Pro Pro Ser Thr Glu Asp Ala 195 200
205Arg Ser Cys Gln Phe Pro Glu Glu Glu Arg Gly Glu Arg Ser Ala Glu
210 215 220Glu Lys Gly Arg Leu Gly Asp Leu Trp Val225
2304137PRTHomo sapiensmisc_feature(1)..(137)extracellular portion
of mature human GITR 4Gln Arg Pro Thr Gly Gly Pro Gly Cys Gly Pro
Gly Arg Leu Leu Leu1 5 10 15Gly Thr Gly Thr Asp Ala Arg Cys Cys Arg
Val His Thr Thr Arg Cys 20 25 30Cys Arg Asp Tyr Pro Gly Glu Glu Cys
Cys Ser Glu Trp Asp Cys Met 35 40 45Cys Val Gln Pro Glu Phe His Cys
Gly Asp Pro Cys Cys Thr Thr Cys 50 55 60Arg His His Pro Cys Pro Pro
Gly Gln Gly Val Gln Ser Gln Gly Lys65 70 75 80Phe Ser Phe Gly Phe
Gln Cys Ile Asp Cys Ala Ser Gly Thr Phe Ser 85 90 95Gly Gly His Glu
Gly His Cys Lys Pro Trp Thr Asp Cys Thr Gln Phe 100 105 110Gly Phe
Leu Thr Val Phe Pro Gly Asn Lys Thr His Asn Ala Val Cys 115 120
125Val Pro Gly Ser Pro Pro Ala Glu Pro 130 13555PRTArtificial
SequenceSynthetic 6G10 VH CDR1 5Thr Tyr Gly Met His1
5617PRTArtificial SequenceSynthetic 6G10 VH CDR2 6Val Thr Trp Tyr
Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val Lys1 5 10
15Gly715PRTArtificial SequenceSynthetic 6G10 VH CDR3 7Gly Gly Ser
Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp Val1 5 10
15811PRTArtificial SequenceSynthetic 6G10 VL CDR1 8Arg Ala Ser Gln
Gly Ile Ser Ser Ala Leu Ala1 5 1097PRTArtificial SequenceSynthetic
6G10 VL CDR2 9Asp Ala Ser Ser Leu Glu Ser1 5109PRTArtificial
SequenceSynthetic 6G10 VL CDR3 10Gln Gln Phe Asn Ser Tyr Pro Tyr
Thr1 511124PRTArtificial SequenceSynthetic 6G10 VH 11Gln Val Gln
Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Thr Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly Leu Tyr Tyr
Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val
Ser Ser 115 12012107PRTArtificial SequenceSynthetic 6G10 VL 12Ala
Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys 100 10513450PRTArtificial SequenceSynthetic 6G10 full length
heavy chain 13Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Thr Trp Tyr Ala Gly Ser Asn Lys
Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met
Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135
140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235 240Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn
Ser Thr Phe Arg 290 295 300Val Val Ser Val Leu Thr Val Val His Gln
Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser
Asn Lys Gly Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys
Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375
380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Met385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys
45014214PRTArtificial SequenceSynthetic 6G10 full length light
chain 14Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser
Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys
Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser
Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln
Gln Phe Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe
Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val
Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155
160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys
21015454PRTArtificial SequenceSynthetic chimeric 6G10 antibodyfull
length heavy chain 15Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val
Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Thr Trp Tyr Ala Gly Ser
Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Lys Thr Thr 115 120
125Ala Pro Ser Val Tyr Pro Leu Ala Pro Val Cys Gly Asp Thr Thr Gly
130 135 140Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Leu Thr Trp Asn Ser Gly Ser Leu Ser
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Asp Leu
Tyr Thr Leu Ser Ser Ser Val 180 185 190Thr Val Thr Ser Ser Thr Trp
Pro Ser Gln Ser Ile Thr Cys Asn Val 195 200 205Ala His Pro Ala Ser
Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg 210 215 220Gly Pro Thr
Ile Lys Pro Cys Pro Pro Cys Lys Cys Pro Ala Pro Asn225 230 235
240Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp
245 250 255Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys Val Val
Val Asp 260 265 270Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp
Phe Val Asn Asn 275 280 285Val Glu Val His Thr Ala Gln Thr Gln Thr
His Arg Glu Asp Tyr Asn 290 295 300Ser Thr Leu Arg Val Val Ser Ala
Leu Pro Ile Gln His Gln Asp Trp305 310 315 320Met Ser Gly Lys Glu
Phe Lys Cys Lys Val Asn Asn Lys Asp Leu Pro 325 330 335Ala Pro Ile
Glu Arg Thr Ile Ser Lys Pro Lys Gly Ser Val Arg Ala 340 345 350Pro
Gln Val Tyr Val Leu Pro Pro Pro Glu Glu Glu Met Thr Lys Lys 355 360
365Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met Pro Glu Asp Ile
370 375 380Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu Asn Tyr
Lys Asn385 390 395 400Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr
Phe Met Tyr Ser Lys 405 410 415Leu Arg Val Glu Lys Lys Asn Trp Val
Glu Arg Asn Ser Tyr Ser Cys 420 425 430Ser Val Val His Glu Gly Leu
His Asn His His Thr Thr Lys Ser Phe 435 440 445Ser Arg Thr Pro Gly
Lys 45016214PRTArtificial SequenceSynthetic chimeric 6G10
antibodyfull length light chain 16Ala Ile Gln Leu Thr Gln Ser Pro
Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys
Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln
Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser
Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly
Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Tyr
85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Ala Asp Ala
Ala 100 105 110Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu
Thr Ser Gly 115 120 125Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe
Tyr Pro Lys Asp Ile 130 135 140Asn Val Lys Trp Lys Ile Asp Gly Ser
Glu Arg Gln Asn Gly Val Leu145 150 155 160Asn Ser Trp Thr Asp Gln
Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser 165 170 175Ser Thr Leu Thr
Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr 180 185 190Thr Cys
Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser 195 200
205Phe Asn Arg Asn Glu Cys 21017372DNAArtificial SequenceSynthetic
6G10 (VH) nucleotide sequence 17caggtgcagc tggtggagtc tgggggagac
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt
acctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg
ggtggcagtt acatggtatg ctggaagtaa taaattttat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc
gagaggaggt 300agtatggttc ggggacttta ttattacggt atggacgtct
ggggccaagg gaccacggtc 360accgtctcct ca 37218321DNAArtificial
SequenceSynthetic 6G10 (VL) nucleotide sequence 18gccatccagt
tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc
gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca
120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg
ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag
tttaatagtt acccgtacac ttttggccag 300gggaccaagc tggagatcaa a
321191350DNAArtificial SequenceSynthetic 6G10 (full length
wild-type heavy chain) nucleotide sequence 19caggtgcagc tggtggagtc
tgggggagac gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt
caccttcagt acctatggca tgcactgggt ccgccaggct 120ccaggcaagg
ggctggagtg ggtggcagtt acatggtatg ctggaagtaa taaattttat
180gcagactccg tgaagggccg attcaccatc tccagagaca attccaagaa
cacgctgtat 240ctgcaaatga acagcctgag agccgaggac acggctgtgt
attactgtgc gagaggaggt 300agtatggttc ggggacttta ttattacggt
atggacgtct ggggccaagg gaccacggtc 360accgtctcct cagcctccac
caagggccca tcggtcttcc ccctggcgcc ctgctccagg 420agcacctccg
agagcacagc ggccctgggc tgcctggtca aggactactt ccccgaaccg
480gtgacggtgt cgtggaactc aggcgctctg accagcggcg tgcacacctt
cccagctgtc 540ctacagtcct caggactcta ctccctcagc agcgtggtga
ccgtgccctc cagcaacttc 600ggcacccaga cctacacctg caacgtagat
cacaagccca gcaacaccaa ggtggacaag 660acagttgagc gcaaatgttg
tgtcgagtgc ccaccgtgcc cagcaccacc tgtggcagga 720ccgtcagtct
tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct
780gaggtcacgt gcgtggtggt ggacgtgagc cacgaagacc ccgaggtcca
gttcaactgg 840tacgtggacg gcgtggaggt gcataatgcc aagacaaagc
cacgggagga gcagttcaac 900agcacgttcc gtgtggtcag cgtcctcacc
gttgtgcacc aggactggct gaacggcaag 960gagtacaagt gcaaggtctc
caacaaaggc ctcccagccc ccatcgagaa aaccatctcc 1020aaaaccaaag
ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggaggag
1080atgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctaccc
cagcgacatc 1140gccgtggagt gggagagcaa tgggcagccg gagaacaact
acaagaccac acctcccatg 1200ctggactccg acggctcctt cttcctctac
agcaagctca ccgtggacaa gagcaggtgg 1260cagcagggga acgtcttctc
atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320cagaagagcc
tctccctgtc tccgggtaaa 135020642DNAArtificial SequenceSynthetic 6G10
(full length wild-type light chain) nucleotide sequence
20gccatccagt tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc
60atcacttgcc gggcaagtca gggcattagc agtgctttag cctggtatca gcagaaacca
120gggaaagctc ctaagctcct gatctatgat gcctccagtt tggaaagtgg
ggtcccatca 180aggttcagcg gcagtggatc tgggacagat ttcactctca
ccatcagcag cctgcagcct 240gaagattttg caacttatta ctgtcaacag
tttaatagtt acccgtacac ttttggccag 300gggaccaagc tggagatcaa
acgaactgtg gctgcaccat ctgtcttcat cttcccgcca 360tctgatgagc
agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat
420cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg
taactcccag 480gagagtgtca cagagcagga cagcaaggac agcacctaca
gcctcagcag caccctgacg 540ctgagcaaag cagactacga gaaacacaaa
gtctacgcct gcgaagtcac ccatcagggc 600ctgagctcgc ccgtcacaaa
gagcttcaac aggggagagt gt 642211365DNAArtificial SequenceSynthetic
chimeric 6G10 heavy chain 21caggtgcagc tggtggagtc tgggggagac
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt
acctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg
ggtggcagtt acatggtatg ctggaagtaa taaattttat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc
gagaggaggt 300agtatggttc ggggacttta ttattacggt atggacgtct
ggggccaagg gaccacggtc 360accgtctcct cagccaaaac aacagcccca
tcggtctatc cgctagcccc tgtgtgtgga 420gatacaactg gctcctcggt
gactctagga tgcctggtca agggttattt ccctgagcca 480gtgaccttga
cctggaactc tggatccctg tccagtggtg tgcacacctt cccagctgtc
540ctgcagtctg acctctacac cctcagcagc tcagtgactg taacctcgag
cacctggccc 600agccagtcca tcacctgcaa tgtggcccac ccggcaagca
gcaccaaggt ggacaagaaa 660attgagccca gagggcccac aatcaagccc
tgtcctccat gcaaatgccc agcacctaac 720ctcttgggtg gaccatccgt
cttcatcttc cctccaaaga tcaaggatgt actcatgatc 780tccctgagcc
ccatagtcac atgtgtggtg gtggatgtga gcgaggatga cccagatgtc
840cagatcagct ggtttgtgaa caacgtggaa gtacacacag ctcagacaca
aacccataga 900gaggattaca acagtactct ccgggtggtc agtgccctcc
ccatccagca ccaggactgg 960atgagtggca aggagttcaa atgcaaggtc
aacaacaaag acctcccagc gcccatcgag 1020agaaccatct caaaacccaa
agggtcagta agagctccac aggtatatgt cttgcctcca 1080ccagaagaag
agatgactaa gaaacaggtc actctgacct gcatggtcac agacttcatg
1140cctgaagaca tttacgtgga gtggaccaac aacgggaaaa cagagctaaa
ctacaagaac 1200actgaaccag tcctggactc tgatggttct tacttcatgt
acagcaagct gagagtggaa 1260aagaagaact gggtggaaag aaatagctac
tcctgttcag tggtccacga gggtctgcac 1320aatcaccaca cgactaagag
cttctcccgg actccgggta aatga 136522645DNAArtificial
SequenceSynthetic chimeric 6G10 light chain 22gccatccagt tgacccagtc
tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca
gggcattagc agtgctttag cctggtatca gcagaaacca 120gggaaagctc
ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca
180aggttcagcg gcagtggatc tgggacagat ttcactctca ccatcagcag
cctgcagcct 240gaagattttg caacttatta ctgtcaacag tttaatagtt
acccgtacac ttttggccag 300gggaccaagc tggagatcaa acgggctgat
gctgcaccaa ctgtatccat cttcccacca 360tccagtgagc aattgacatc
tggaggtgcc tcagtcgtgt gcttcttgaa caacttctac 420cccaaagaca
tcaatgtcaa gtggaagatt gatggcagtg aacgacaaaa tggcgtcctg
480aacagttgga ccgatcagga cagcaaagac agcacctaca gcatgagcag
caccctcacg 540ttgaccaagg acgagtatga acgacataac agctatacct
gtgaggccac tcacaagaca 600tcaacttcac ccattgtcaa gagcttcaac
aggaatgagt gttag 64523326PRTHomo sapiensmisc_feature(1)..(326)human
IgG2 heavy chain constant region 23Ala Ser Thr Lys Gly Pro Ser Val
Phe Pro Leu Ala Pro Cys Ser Arg1 5 10 15Ser Thr Ser Glu Ser Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr 20 25 30Phe Pro Glu Pro Val Thr
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 35 40 45Gly Val His Thr Phe
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser 50 55 60Leu Ser Ser Val
Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr65 70 75 80Tyr Thr
Cys Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95Thr
Val Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro 100 105
110Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
115 120 125Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp 130 135 140Val Ser His Glu Asp Pro Glu Val Gln Phe Asn Trp
Tyr Val Asp Gly145 150 155 160Val Glu Val His Asn Ala Lys Thr Lys
Pro Arg Glu Glu Gln Phe Asn 165 170 175Ser Thr Phe Arg Val Val Ser
Val Leu Thr Val Val His Gln Asp Trp 180 185 190Leu Asn Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro 195 200 205Ala Pro Ile
Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu 210 215 220Pro
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn225 230
235 240Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile 245 250 255Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr 260 265 270Thr Pro Pro Met Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys 275 280 285Leu Thr Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys 290 295 300Ser Val Met His Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu305 310 315 320Ser Leu Ser Pro
Gly Lys 32524107PRTHomo sapiensmisc_feature(1)..(107)human kappa
light chain constant region 24Arg Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu1 5 10 15Gln Leu Lys Ser Gly Thr Ala Ser
Val Val Cys Leu Leu Asn Asn Phe 20 25 30Tyr Pro Arg Glu Ala Lys Val
Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45Ser Gly Asn Ser Gln Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60Thr Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu65 70 75 80Lys His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95Pro Val
Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 10525330PRTMus
musculusmisc_feature(1)..(330)mouse IgG2a heavy chain constant
region 25Ala Lys Thr Thr Ala Pro Ser Val Tyr Pro Leu Ala Pro Val
Cys Gly1 5 10 15Asp Thr Thr Gly Ser Ser Val Thr Leu Gly Cys Leu Val
Lys Gly Tyr 20 25 30Phe Pro Glu Pro Val Thr Leu Thr Trp Asn Ser Gly
Ser Leu Ser Ser 35 40 45Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
Asp Leu Tyr Thr Leu 50 55 60Ser Ser Ser Val Thr Val Thr Ser Ser Thr
Trp Pro Ser Gln Ser Ile65 70 75 80Thr Cys Asn Val Ala His Pro Ala
Ser Ser Thr Lys Val Asp Lys Lys 85 90 95Ile Glu Pro Arg Gly Pro Thr
Ile Lys Pro Cys Pro Pro Cys Lys Cys 100 105 110Pro Ala Pro Asn Leu
Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro 115 120 125Lys Ile Lys
Asp Val Leu Met Ile Ser Leu Ser Pro Ile Val Thr Cys 130 135 140Val
Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp145 150
155 160Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His
Arg 165 170 175Glu Asp Tyr Asn Ser Thr Leu Arg Val Val Ser Ala Leu
Pro Ile Gln 180 185 190His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys
Cys Lys Val Asn Asn 195 200 205Lys Asp Leu Pro Ala Pro Ile Glu Arg
Thr Ile Ser Lys Pro Lys Gly 210 215 220Ser Val Arg Ala Pro Gln Val
Tyr Val Leu Pro Pro Pro Glu Glu Glu225 230 235 240Met Thr Lys Lys
Gln Val Thr Leu Thr Cys Met Val Thr Asp Phe Met 245 250 255Pro Glu
Asp Ile Tyr Val Glu Trp Thr Asn Asn Gly Lys Thr Glu Leu 260 265
270Asn Tyr Lys Asn Thr Glu Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe
275 280 285Met Tyr Ser Lys Leu Arg Val Glu Lys Lys Asn Trp Val Glu
Arg Asn 290 295 300Ser Tyr Ser Cys Ser Val Val His Glu Gly Leu His
Asn His His Thr305 310 315 320Thr Lys Ser Phe Ser Arg Thr Pro Gly
Lys 325 33026107PRTMus musculusmisc_feature(1)..(107)mouse kappa
light chain constant region 26Arg Ala Asp Ala Ala Pro Thr Val Ser
Ile Phe Pro Pro Ser Ser Glu1 5 10 15Gln Leu Thr Ser Gly Gly Ala Ser
Val Val Cys Phe Leu Asn Asn Phe 20 25 30Tyr Pro Lys Asp Ile Asn Val
Lys Trp Lys Ile Asp Gly Ser Glu Arg 35 40 45Gln Asn Gly Val Leu Asn
Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser 50 55 60Thr Tyr Ser Met Ser
Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu65 70 75 80Arg His Asn
Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser 85 90 95Pro Ile
Val Lys Ser Phe Asn Arg Asn Glu Cys 100 10527124PRTArtificial
SequenceSynthetic 28F3 (VH) 27Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu
Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Ser Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115
12028107PRTArtificial SequenceSynthetic 28F3 (VL) 28Ala Ile Gln Leu
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr
Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro
Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
10529450PRTArtificial SequenceSynthetic 28F3 (full length wild-type
heavy chain) 29Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met
Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135
140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235 240Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn
Ser Thr Phe Arg 290 295 300Val Val Ser
Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly Lys305 310 315
320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu
325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln
Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn
Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn
Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu Asp Ser Asp Gly
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly Lys 45030214PRTArtificial SequenceSynthetic 28F3 (full
length wild-type light chain) 30Ala Ile Gln Leu Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu
Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Tyr 85 90 95Thr Phe
Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105
110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly
115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg
Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser
Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys
Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys
Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr
His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg
Gly Glu Cys 21031453PRTArtificial SequenceSynthetic 28F3.IgG1 (VH +
IgG1) 31Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly
Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu
Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr
Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser
Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu
Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg
Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr
Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Leu Leu Gly Gly Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
45032453PRTArtificial SequenceSynthetic 28F3.IgG1.1 (VH + IgG1.1)
32Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly
Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
45033214PRTArtificial SequenceSynthetic 28F3.IgG1 (VL + CL) 33Ala
Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 210345PRTArtificial
SequenceSynthetic 28F3 VH CDR1 34Ser Tyr Gly Met His1
53517PRTArtificial SequenceSynthetic 28F3 VH CDR2 35Val Ile Trp Tyr
Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10
15Gly3615PRTArtificial SequenceSynthetic 28F3 VH CDR3 36Gly Gly Ser
Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp Val1 5 10
153711PRTArtificial SequenceSynthetic 28F3 VL CDR1 37Arg Ala Ser
Gln Gly Ile Ser Ser Ala Leu Ala1 5 10387PRTArtificial
SequenceSynthetic 28F3 VL CDR2 38Asp Ala Ser Ser Leu Glu Ser1
5399PRTArtificial SequenceSynthetic 28F3 VL CDR3 39Gln Gln Phe Asn
Ser Tyr Pro Tyr Thr1 540449PRTArtificial SequenceSynthetic 28F3 (VH
+ G2(C219S)) or 28F3-IgG2- C219S 40Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr
Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Ser Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230
235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Phe Asn Ser Thr Phe Arg 290 295 300Val Val Ser Val Leu Thr Val
Val His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr
Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Met385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly41450PRTArtificial SequenceSynthetic 28F3 (VH + G2.g1) or
28F3-IgG2-IgG1 41Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn
Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser
Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135
140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235 240Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250
255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys Thr Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr Thr Leu
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375
380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro
Gly 45042449PRTArtificial SequenceSynthetic 28F3 (VH + G2.g1.1) or
28F3-IgG2- IgG1.1 42Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn
Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser
Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135
140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235 240Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375
380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly43450PRTArtificial
SequenceSynthetic 28F3 (VH + G2(C219S).g1) or 28F3- IgG2-C219S-IgG1
43Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly
Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr
Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val
Asp Lys Thr Val Glu Arg 210 215 220Lys Ser Cys Val Glu Cys Pro Pro
Cys Pro Ala Pro Glu Leu Leu Gly225 230 235 240Gly Pro Ser Val Phe
Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250 255Ile Ser Arg
Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 260 265 270Glu
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280
285His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr
290 295 300Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu
Asn Gly305 310 315 320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala
Leu Pro Ala Pro Ile 325 330 335Glu Lys Thr Ile Ser Lys Ala Lys Gly
Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg
Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395
400Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val
405 410 415Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser
Val Met 420 425 430His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser 435 440 445Pro Gly 45044449PRTArtificial
SequenceSynthetic 28F3 (VH + G2(C219S).g1.1) or 28F3-
IgG2-C219S-IgG1.1 44Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn
Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser
Met Val Arg Gly Asp Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly
Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135
140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Ser Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235 240Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250
255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375
380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly45141PRTArtificial
SequenceSynthetic 28F3 (VH) (SEQ ID NO 13) with signal peptide
45Met Arg Ala Trp Ile Phe Phe Leu Leu Cys Leu Ala Gly Arg Ala Leu1
5 10 15Ala Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
Gly 20 25 30Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Ser Ser 35 40 45Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp 50 55 60Val Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr
Tyr Ala Asp Ser65 70 75 80Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu 85 90 95Tyr Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr 100 105 110Cys Ala Arg Gly Gly Ser Met
Val Arg Gly Asp Tyr Tyr Tyr Gly Met 115 120 125Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser 130 135 14046124PRTArtificial
SequenceSynthetic 28F3 (VL) (SEQ ID NO 14) with signal peptide
46Met Arg Ala Trp Ile Phe Phe Leu Leu Cys Leu Ala Gly Arg Ala Leu1
5 10 15Ala Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser
Val 20 25 30Gly Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile
Ser Ser 35 40 45Ala Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro
Lys Leu Leu 50 55 60Ile Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro
Ser Arg Phe Ser65 70 75 80Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln 85 90 95Pro Glu Asp Phe Ala Thr Tyr Tyr Cys
Gln Gln Phe Asn Ser Tyr Pro 100 105 110Tyr Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 115 12047470PRTArtificial SequenceSynthetic
28F3.IgG1 (VH + IgG1) with signal peptide 47Met Arg Ala Trp Ile Phe
Phe Leu Leu Cys Leu Ala Gly Arg Ala Leu1 5 10 15Ala Gln Val Gln Leu
Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly 20 25 30Arg Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 35 40 45Tyr Gly Met
His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 50 55 60Val Ala
Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser65 70 75
80Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
85 90 95Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr
Tyr 100 105 110Cys Ala Arg Gly Gly Ser Met Val Arg Gly Asp Tyr Tyr
Tyr Gly Met 115 120 125Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val
Ser Ser Ala Ser Thr 130 135 140Lys Gly Pro Ser Val Phe Pro Leu Ala
Pro Ser Ser Lys Ser Thr Ser145 150 155 160Gly Gly Thr Ala Ala Leu
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 165 170 175Pro Val Thr Val
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 180 185 190Thr Phe
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 195 200
205Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
210 215 220Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg
Val Glu225 230 235 240Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
Pro Cys Pro Ala Pro 245 250 255Glu Leu Leu Gly Gly Pro Ser Val Phe
Leu Phe Pro Pro Lys Pro Lys 260 265 270Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val Thr Cys Val Val Val 275 280 285Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 290 295 300Gly Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr305 310 315
320Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
325 330 335Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys
Ala Leu 340 345 350Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys
Gly Gln Pro Arg 355 360 365Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
Arg Glu Glu Met Thr Lys 370 375 380Asn Gln Val Ser Leu Thr Cys Leu
Val Lys Gly Phe Tyr Pro Ser Asp385 390 395 400Ile Ala Val Glu Trp
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 405 410 415Thr Thr Pro
Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430Lys
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440
445Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
450 455 460Leu Ser Leu Ser Pro Gly465 47048470PRTArtificial
SequenceSynthetic 28F3.IgG1.1 (VH + IgG1.1) with signal peptide
48Met Arg Ala Trp Ile Phe Phe Leu Leu Cys Leu Ala Gly Arg Ala Leu1
5 10 15Ala Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro
Gly 20 25 30Arg Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Ser Ser 35 40 45Tyr Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp 50 55 60Val Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr
Tyr Ala Asp Ser65 70 75 80Val Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu 85 90 95Tyr Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr 100 105 110Cys Ala Arg Gly Gly Ser Met
Val Arg Gly Asp Tyr Tyr Tyr Gly Met 115 120 125Asp Val Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr 130 135 140Lys Gly Pro
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser145 150 155
160Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
165 170 175Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His 180 185 190Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser 195 200 205Val Val Thr Val Pro Ser Ser Ser Leu Gly
Thr Gln Thr Tyr Ile Cys 210 215 220Asn Val Asn His Lys Pro Ser Asn
Thr Lys Val Asp Lys Arg Val Glu225 230 235 240Pro Lys Ser Cys Asp
Lys Thr His Thr Cys Pro Pro Cys Pro Ala
Pro 245 250 255Glu Ala Glu Gly Ala Pro Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys 260 265 270Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val Val 275 280 285Asp Val Ser His Glu Asp Pro Glu Val
Lys Phe Asn Trp Tyr Val Asp 290 295 300Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln Tyr305 310 315 320Asn Ser Thr Tyr
Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 325 330 335Trp Leu
Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 340 345
350Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
355 360 365Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
Thr Lys 370 375 380Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp385 390 395 400Ile Ala Val Glu Trp Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys 405 410 415Thr Thr Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser 420 425 430Lys Leu Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 435 440 445Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 450 455 460Leu
Ser Leu Ser Pro Gly465 47049231PRTArtificial SequenceSynthetic
28F3.IgG1 (VL+CL) with signal peptide 49Met Arg Ala Trp Ile Phe Phe
Leu Leu Cys Leu Ala Gly Arg Ala Leu1 5 10 15Ala Ala Ile Gln Leu Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val 20 25 30Gly Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser 35 40 45Ala Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu 50 55 60Ile Tyr Asp
Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser65 70 75 80Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln 85 90
95Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro
100 105 110Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr
Val Ala 115 120 125Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
Gln Leu Lys Ser 130 135 140Gly Thr Ala Ser Val Val Cys Leu Leu Asn
Asn Phe Tyr Pro Arg Glu145 150 155 160Ala Lys Val Gln Trp Lys Val
Asp Asn Ala Leu Gln Ser Gly Asn Ser 165 170 175Gln Glu Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 180 185 190Ser Ser Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 195 200 205Tyr
Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 210 215
220Ser Phe Asn Arg Gly Glu Cys225 23050123PRTArtificial
SequenceSynthetic 19D3 (VH) 50Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala
Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105
110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115
12051107PRTArtificial SequenceSynthetic 19D3 (VL) 51Asp Ile Gln Met
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr
Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro
Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
10552449PRTArtificial SequenceSynthetic 19D3 (full length wild-type
heavy chain) 52Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys
Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Gln Leu
Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp Gly Gln Gly
Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125Pro Ser
Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 130 135
140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe Gly
Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser Asn
Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Cys Cys Val Glu
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Phe Arg Val 290 295 300Val Ser Val Leu Thr Val Val His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ala Pro Ile Glu Lys 325 330 335Thr Ile Ser Lys Thr
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met
Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445Lys53214PRTArtificial
SequenceSynthetic 19D3 (full length wild-type light chain) 53Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu
Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 21054452PRTArtificial
SequenceSynthetic 19D3.IgG1 (VH + IgG1) 54Gln Val Gln Leu Val Glu
Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile
Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp
Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200
205Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys
210 215 220Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
Glu Leu225 230 235 240Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr 245 250 255Leu Met Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val Val Asp Val 260 265 270Ser His Glu Asp Pro Glu Val
Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280 285Glu Val His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290 295 300Thr Tyr Arg
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu305 310 315
320Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala
325 330 335Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro 340 345 350Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
Thr Lys Asn Gln 355 360 365Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala 370 375 380Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr385 390 395 400Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410 415Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 420 425 430Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440
445Leu Ser Pro Gly 45055452PRTArtificial SequenceSynthetic
19D3.IgG1.1 (VH + IgG1.1) 55Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly
Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120
125Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly
130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Ser Leu
Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200 205Asn His Lys Pro Ser
Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys 210 215 220Ser Cys Asp
Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala225 230 235
240Glu Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
245 250 255Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val 260 265 270Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val 275 280 285Glu Val His Asn Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser 290 295 300Thr Tyr Arg Val Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu305 310 315 320Asn Gly Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser 325 330 335Ser Ile Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 340 345 350Gln
Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln 355 360
365Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
370 375 380Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr385 390 395 400Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu 405 410 415Thr Val Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser 420 425 430Val Met His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440 445Leu Ser Pro Gly
45056214PRTArtificial SequenceSynthetic 19D3.IgG1 (VL + CL) 56Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu
Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser
Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 210575PRTArtificial
SequenceSynthetic 19D3 VH CDR1 57Ser Tyr Gly Phe His1
55817PRTArtificial SequenceSynthetic 19D3 VH CDR2 58Val Ile Trp Tyr
Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val Lys1 5 10
15Gly5914PRTArtificial SequenceSynthetic 19D3 VH CDR3 59Gly Gly Gln
Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val1 5 106011PRTArtificial
SequenceSynthetic 19D3 VL CDR1 60Arg Ala Ser Gln Gly Ile Ser Ser
Trp Leu Ala1 5 10617PRTArtificial SequenceSynthetic 19D3 VL CDR2
61Ala Ala Ser Ser Leu Gln Ser1 5629PRTArtificial SequenceSynthetic
19D3 VL CDR3 62Gln Gln Tyr Asn Ser Tyr Pro Tyr Thr1
563448PRTArtificial SequenceSynthetic 19D3 (VH + G2(C219S)) or
19D3-IgG2- C219S 63Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser Asn
Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Gln
Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125Pro
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 130 135
140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe Gly
Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser Asn
Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Ser Cys Val Glu
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Phe Arg Val 290 295 300Val Ser Val Leu Thr Val Val His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ala Pro Ile Glu Lys 325 330 335Thr Ile Ser Lys Thr
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met
Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly 435 440 44564449PRTArtificial
SequenceSynthetic 19D3 (VH + G2.g1) or 19D3-IgG2-IgG1 64Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly
Phe His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr
Val Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser
Ser Ala Ser Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro
Cys Ser Arg Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185
190Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val
195 200 205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu
Arg Lys 210 215 220Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu
Leu Leu Gly Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn Ala Lys
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys305 310
315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu 325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425
430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445Gly65448PRTArtificial SequenceSynthetic 19D3 (VH +
G2.g1.1) or 19D3-IgG2- IgG1.1 65Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala
Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105
110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser
Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu
Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser
Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser
Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys
Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Cys
Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230
235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345
350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44566449PRTArtificial SequenceSynthetic 19D3 (VH + G2(C219S).g1) or
19D3- IgG2-C219S-IgG1 66Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser
Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120
125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Ser Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly67448PRTArtificial SequenceSynthetic 19D3 (VH +
G2(C219S).g1.1) or 19D3- IgG2-C219S-IgG1.1 67Gln Val Gln Leu Val
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val
Ile Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp
Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val
Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200
205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys
210 215 220Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala
Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315
320Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys
325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr 340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44568448PRTArtificial SequenceSynthetic 19D3 (VH + G2(C219S))
or
19D3-IgG2- C219S 68Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser Asn
Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu
Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Gln
Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp Gly Gln
Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125Pro
Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 130 135
140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly
Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe Gly
Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser Asn
Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Ser Cys Val Glu
Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230 235 240Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250
255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp
260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val
His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser
Thr Phe Arg Val 290 295 300Val Ser Val Leu Thr Val Val His Gln Asp
Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val Ser Asn
Lys Gly Leu Pro Ala Pro Ile Glu Lys 325 330 335Thr Ile Ser Lys Thr
Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu Pro Pro
Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360 365Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375
380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met
Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly 435 440 44569449PRTArtificial
SequenceSynthetic 19D3 (VH + G2.g1) or 19D3-IgG2-IgG1 69Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly
Phe His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr
Val Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser
Ser Ala Ser Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro
Cys Ser Arg Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser
Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro
Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185
190Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val
195 200 205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu
Arg Lys 210 215 220Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu
Leu Leu Gly Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys
Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr
Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu Val Lys
Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn Ala Lys
Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val
Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys305 310
315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu 325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425
430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
435 440 445Gly70448PRTArtificial SequenceSynthetic 19D3 (VH +
G2.g1.1) or 19D3-IgG2- IgG1.1 70Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala
Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105
110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly
115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser
Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu
Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser
Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser
Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys
Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Cys
Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230
235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys 325 330 335Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345
350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr
355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44571449PRTArtificial SequenceSynthetic 19D3 (VH + G2(C219S).g1) or
19D3- IgG2-C219S-IgG1 71Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser
Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp Val 100 105 110Trp Gly
Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120
125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Ser Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly72448PRTArtificial SequenceSynthetic 19D3 (VH +
G2(C219S).g1.1) or 19D3- IgG2-C219S-IgG1.1 72Gln Val Gln Leu Val
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Phe His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val
Ile Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Gln Leu Asp Tyr Tyr Tyr Tyr Tyr Val Met Asp
Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val
Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200
205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys
210 215 220Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala
Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315
320Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys
325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr 340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44573123PRTArtificial SequenceSynthetic 18E10 (VH) 73Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly
Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe Tyr Tyr
Ser Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser
Ser 115 12074107PRTArtificial SequenceSynthetic 18E10 (VL) 74Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu
Ile
35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu
Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn
Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
100 10575449PRTArtificial SequenceSynthetic 18E10 (full length
wild-type heavy chain) 75Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly
Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp Val 100 105 110Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120
125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Cys Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly Pro225 230 235
240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser
245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
Glu Asp 260 265 270Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val
Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe
Asn Ser Thr Phe Arg Val 290 295 300Val Ser Val Leu Thr Val Val His
Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr Lys Cys Lys Val
Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys 325 330 335Thr Ile Ser
Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350Leu
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr 355 360
365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Met Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
445Lys76214PRTArtificial SequenceSynthetic 18E10 (full length
wild-type light chain) 76Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala
Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro
Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln
Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly
Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120
125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu
Cys 21077452PRTArtificial SequenceSynthetic 18E10.IgG1 (VH + IgG1)
77Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Ile Ala Val Ala
Phe Tyr Tyr Ser Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val
Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120 125Pro Ser Val Phe Pro
Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly 130 135 140Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val145 150 155
160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe
165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
Val Val 180 185 190Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr
Ile Cys Asn Val 195 200 205Asn His Lys Pro Ser Asn Thr Lys Val Asp
Lys Arg Val Glu Pro Lys 210 215 220Ser Cys Asp Lys Thr His Thr Cys
Pro Pro Cys Pro Ala Pro Glu Leu225 230 235 240Leu Gly Gly Pro Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 245 250 255Leu Met Ile
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 260 265 270Ser
His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280
285Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser
290 295 300Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
Trp Leu305 310 315 320Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn
Lys Ala Leu Pro Ala 325 330 335Pro Ile Glu Lys Thr Ile Ser Lys Ala
Lys Gly Gln Pro Arg Glu Pro 340 345 350Gln Val Tyr Thr Leu Pro Pro
Ser Arg Glu Glu Met Thr Lys Asn Gln 355 360 365Val Ser Leu Thr Cys
Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 370 375 380Val Glu Trp
Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr385 390 395
400Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu
405 410 415Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
Cys Ser 420 425 430Val Met His Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser 435 440 445Leu Ser Pro Gly 45078452PRTArtificial
SequenceSynthetic 18E10.IgG1.1 (VH + IgG1.1) 78Gln Val Gln Leu Val
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val
Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp
Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val
Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val 195 200
205Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys
210 215 220Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
Glu Ala225 230 235 240Glu Gly Ala Pro Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr 245 250 255Leu Met Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val Val Asp Val 260 265 270Ser His Glu Asp Pro Glu Val
Lys Phe Asn Trp Tyr Val Asp Gly Val 275 280 285Glu Val His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 290 295 300Thr Tyr Arg
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu305 310 315
320Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser
325 330 335Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
Glu Pro 340 345 350Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met
Thr Lys Asn Gln 355 360 365Val Ser Leu Thr Cys Leu Val Lys Gly Phe
Tyr Pro Ser Asp Ile Ala 370 375 380Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu Asn Asn Tyr Lys Thr Thr385 390 395 400Pro Pro Val Leu Asp
Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 405 410 415Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 420 425 430Val
Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 435 440
445Leu Ser Pro Gly 45079214PRTArtificial SequenceSynthetic
18E10.IgG1 (VL + CL) 79Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu
Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu
Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln
Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120
125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu
Cys 210805PRTArtificial SequenceSynthetic 18E10 VH CDR1 80Ser Tyr
Gly Met His1 58117PRTArtificial SequenceSynthetic 18E10 VH CDR2
81Val Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys1
5 10 15Gly8214PRTArtificial SequenceSynthetic 18E10 VH CDR3 82Gly
Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp Val1 5
108311PRTArtificial SequenceSynthetic 18E10 VL CDR1 83Arg Ala Ser
Gln Gly Ile Ser Ser Trp Leu Ala1 5 10847PRTArtificial
SequenceSynthetic 18E10 VL CDR2 84Ala Ala Ser Ser Leu Gln Ser1
5859PRTArtificial SequenceSynthetic 18E10 VL CDR3 85Gln Gln Tyr Asn
Ser Tyr Pro Tyr Thr1 586448PRTArtificial SequenceSynthetic 18E10
(VH + G2(C219S)) or 18E10- IgG2-C219S 86Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp
Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp Val
100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro
Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp
His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215
220Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly
Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Gln Phe Asn Trp Tyr
Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg
Glu Glu Gln Phe Asn Ser Thr Phe Arg Val 290 295 300Val Ser Val Leu
Thr Val Val His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr
Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys 325 330
335Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro Met Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44587449PRTArtificial SequenceSynthetic 18E10 (VH + G2.g1) or
18E10-IgG2- IgG1 87Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val
Gln Pro Gly Arg1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ala Val Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe
Tyr Tyr Ser Met Asp Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr
Val Ser Ser Ala Ser Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu
Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170
175Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val
180 185 190Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys
Asn Val 195 200 205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr
Val Glu Arg Lys 210 215 220Cys Cys Val Glu Cys Pro Pro Cys Pro Ala
Pro Glu Leu Leu Gly Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295
300Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly88448PRTArtificial SequenceSynthetic 18E10
(VH + G2.g1.1) or 18E10-IgG2- IgG1.1 88Gln Val Gln Leu Val Glu Ser
Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp
Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90
95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp Val
100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr
Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro
Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp
His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215
220Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly
Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315 320Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys 325 330
335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44589449PRTArtificial SequenceSynthetic 18E10 (VH + G2(C219S).g1)
or 18E10- IgG2-C219S-IgG1 89Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Ala Gly
Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp Val 100 105 110Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly 115 120
125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val145 150 155 160Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser
Gly Val His Thr Phe 165 170 175Pro Ala Val Leu Gln Ser Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val Pro Ser Ser Asn Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200 205Asp His Lys Pro Ser
Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys 210 215 220Ser Cys Val
Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly90448PRTArtificial SequenceSynthetic 18E10 (VH +
G2(C219S).g1.1) or 18E10-IgG2-C219S-IgG1.1 90Gln Val Gln Leu Val
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val
Ile Trp Tyr Ala Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Arg Ile Ala Val Ala Phe Tyr Tyr Ser Met Asp
Val 100 105 110Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser
Thr Lys Gly 115 120 125Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg
Ser Thr Ser Glu Ser 130 135 140Thr Ala Ala Leu Gly Cys Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val145 150 155 160Thr Val Ser Trp Asn Ser
Gly Ala Leu Thr Ser Gly Val His Thr Phe 165 170 175Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val 180 185 190Thr Val
Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val 195 200
205Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg Lys
210 215 220Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala
Gly Pro225 230 235 240Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser 245 250 255Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp Val Ser His Glu Asp 260 265 270Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly Val Glu Val His Asn 275 280 285Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300Val Ser Val
Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu305 310 315
320Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys
325 330 335Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val
Tyr Thr 340 345 350Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln
Val Ser Leu Thr 355 360 365Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
Ile Ala Val Glu Trp Glu 370 375 380Ser Asn Gly Gln Pro Glu Asn Asn
Tyr Lys Thr Thr Pro Pro Val Leu385 390 395 400Asp Ser Asp Gly Ser
Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415Ser Arg Trp
Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430Ala
Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440
44591117PRTArtificial SequenceSynthetic 3C3 (VH) 91Gln Val Gln Leu
Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser
Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp
Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly
Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 50 55
60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65
70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp Gly Gln Gly
Thr Met 100 105 110Val Thr Val Ser Ser 11592107PRTArtificial
SequenceSynthetic 3C3 (VL1) 92Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe
Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10593106PRTArtificial
SequenceSynthetic 3C3 (VL2) 93Glu Ile Val Leu Thr Gln Ser Pro Ala
Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg
Ala Ser Gln Gly Val Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg
Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Pro Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe
Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp His Thr 85 90 95Phe Gly
Gln Gly Thr Lys Leu Glu Ile Lys 100 10594447PRTArtificial
SequenceSynthetic 3C3 (full length wild-type heavy chain) 94Gln Val
Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr
Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25
30Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45Gly Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu
Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val
Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp
Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Ser Ser Lys Ser Thr
Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170
175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
Ser Asn 195 200 205Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys
Asp Lys Thr His 210 215 220Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
Leu Gly Gly Pro Ser Val225 230 235 240Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285Thr
Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys305 310 315 320Cys Lys
Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile 325 330
335Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro
340 345 350Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr
Cys Leu 355 360 365Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
Trp Glu Ser Asn 370 375 380Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro Val Leu Asp Ser385 390 395 400Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440
44595214PRTArtificial SequenceSynthetic 3C3 L1 (full length
wild-type light chain 1) 95Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala
Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro
Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln
Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly
Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120
125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu
Cys 21096213PRTArtificial SequenceSynthetic 3C3 L2 (full length
wild-type light chain 2) 96Glu Ile Val Leu Thr Gln Ser Pro Ala Thr
Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala
Ser Gln Gly Val Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala
Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Pro Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala
Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp His Thr 85 90 95Phe Gly Gln
Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110Ser
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120
125Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys
130 135 140Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
Gln Glu145 150 155 160Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
Tyr Ser Leu Ser Ser 165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr
Glu Lys His Lys Val Tyr Ala 180 185 190Cys Glu Val Thr His Gln Gly
Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205Asn Arg Gly Glu Cys
21097446PRTArtificial SequenceSynthetic 3C3.IgG1 (VH + IgG1) 97Gln
Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10
15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr
20 25 30Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
Ile 35 40 45Gly Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser
Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln
Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile
Trp Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr
Lys Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Ser Ser Lys Ser
Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170
175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser
180 185 190Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro
Ser Asn 195 200 205Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys
Asp Lys Thr His 210 215 220Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu
Leu Gly Gly Pro Ser Val225 230 235 240Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280 285Thr
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295
300Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr
Lys305 310 315 320Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
Glu Lys Thr Ile 325 330 335Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val Tyr Thr Leu Pro 340 345 350Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser Leu Thr Cys Leu 355 360 365Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser385 390 395 400Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410
415Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu
420 425 430His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly
435 440 44598446PRTArtificial SequenceSynthetic 3C3.IgG1.1 (VH +
IgG1.1) 98Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro
Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe
Ser Gly Tyr 20 25 30Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly
Leu Glu Trp Ile 35 40 45Gly Lys Ile Asn His Ser Gly Asn Thr Asn Tyr
Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser
Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala
Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala
Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Ser
Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140Leu
Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150
155 160Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln
Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser 180 185 190Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn 195 200 205Thr Lys Val Asp Lys Arg Val Glu Pro
Lys Ser Cys Asp Lys Thr His 210 215 220Thr Cys Pro Pro Cys Pro Ala
Pro Glu Ala Glu Gly Ala Pro Ser Val225 230 235 240Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265
270Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys
275 280 285Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val
Val Ser 290 295 300Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly
Lys Glu Tyr Lys305 310 315 320Cys Lys Val Ser Asn Lys Ala Leu Pro
Ser Ser Ile Glu Lys Thr Ile 325 330 335Ser Lys Ala Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345 350Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser385 390
395 400Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg 405 410 415Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu 420 425 430His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440 44599214PRTArtificial SequenceSynthetic
3C3.IgG1 (VL1 + CL) 99Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu
Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser
Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu
Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser
Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe
Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr
Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln
Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser
Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120
125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu
Cys 210100213PRTArtificial SequenceSynthetic 3C3IgG1.2 (VL2 + CL)
100Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1
5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Gly Val Ser Ser
Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg
Phe Ser Gly 50 55 60Ser Gly Pro Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
Arg Ser Asn Trp His Thr 85 90 95Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala Pro 100 105 110Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu145 150 155
160Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser
165 170 175Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
Tyr Ala 180 185 190Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
Thr Lys Ser Phe 195 200 205Asn Arg Gly Glu Cys 2101015PRTArtificial
SequenceSynthetic 3C3 VH CDR1 101Gly Tyr Tyr Trp Thr1
510216PRTArtificial SequenceSynthetic 3C3 VH CDR2 102Lys Ile Asn
His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys Ser1 5 10
151039PRTArtificial SequenceSynthetic 3C3 VH CDR3 103Leu Gly Ala
Phe Asp Ala Phe Asp Ile1 510411PRTArtificial SequenceSynthetic 3C3
VL1 CDR1 104Arg Ala Ser Gln Gly Ile Ser Ser Trp Leu Ala1 5
101057PRTArtificial SequenceSynthetic 3C3 VL1 CDR2 105Ala Ala Ser
Ser Leu Gln Ser1 51069PRTArtificial SequenceSynthetic 3C3 VL1 CDR3
106Gln Gln Tyr Asn Ser Tyr Pro Tyr Thr1 510711PRTArtificial
SequenceSynthetic 3C3 VL2 CDR1 107Arg Ala Ser Gln Gly Val Ser Ser
Tyr Leu Ala1 5 101087PRTArtificial SequenceSynthetic 3C3 VL2 CDR2
108Asp Ala Ser Asn Arg Ala Thr1 51098PRTArtificial
SequenceSynthetic 3C3 VL2 CDR3 109Gln Gln Arg Ser Asn Trp His Thr1
5110442PRTArtificial SequenceSynthetic 3C3 (VH + G2) or 3C3-IgG2
110Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1
5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly
Tyr 20 25 30Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
Trp Ile 35 40 45Gly Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro
Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn
Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr
Ala Val Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp
Ile Trp Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser
Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Cys Ser Arg
Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155
160Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
Ser Asn 180 185 190Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His
Lys Pro Ser Asn 195 200 205Thr Lys Val Asp Lys Thr Val Glu Arg Lys
Cys Cys Val Glu Cys Pro 210 215 220Pro Cys Pro Ala Pro Pro Val Ala
Gly Pro Ser Val Phe Leu Phe Pro225 230 235 240Pro Lys Pro Lys Asp
Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr 245 250 255Cys Val Val
Val Asp Val Ser His Glu Asp Pro Glu Val Gln Phe Asn 260 265 270Trp
Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 275 280
285Glu Glu Gln Phe Asn Ser Thr Phe Arg Val Val Ser Val Leu Thr Val
290 295 300Val His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
Val Ser305 310 315 320Asn Lys Gly Leu Pro Ala Pro Ile Glu Lys Thr
Ile Ser Lys Thr Lys 325 330 335Gly Gln Pro Arg Glu Pro Gln Val Tyr
Thr Leu Pro Pro Ser Arg Glu 340 345 350Glu Met Thr Lys Asn Gln Val
Ser Leu Thr Cys Leu Val Lys Gly Phe 355 360 365Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 370 375
380Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly Ser
Phe385 390 395 400Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
Trp Gln Gln Gly 405 410 415Asn Val Phe Ser Cys Ser Val Met His Glu
Ala Leu His Asn His Tyr 420 425 430Thr Gln Lys Ser Leu Ser Leu Ser
Pro Gly 435 440111442PRTArtificial SequenceSynthetic 3C3 (VH +
G2(C219S)) or 3C3-IgG2- C219S 111Gln Val Gln Leu Gln Gln Trp Gly
Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala
Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Thr Trp Ile Arg
Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Lys Ile Asn His
Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr
Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu
Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg
Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105
110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr Gln Thr
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr Lys Val
Asp Lys Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro 210 215 220Pro
Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro225 230
235 240Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr 245 250 255Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
Gln Phe Asn 260 265 270Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro Arg 275 280 285Glu Glu Gln Phe Asn Ser Thr Phe Arg
Val Val Ser Val Leu Thr Val 290 295 300Val His Gln Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val Ser305 310 315 320Asn Lys Gly Leu
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys 325 330 335Gly Gln
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 340 345
350Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
355 360 365Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu 370 375 380Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser
Asp Gly Ser Phe385 390 395 400Phe Leu Tyr Ser Lys Leu Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly 405 410 415Asn Val Phe Ser Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr 420 425 430Thr Gln Lys Ser Leu
Ser Leu Ser Pro Gly 435 440112443PRTArtificial SequenceSynthetic
3C3 (VH + G2.g1) or 3C3-IgG2-IgG1 112Gln Val Gln Leu Gln Gln Trp
Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys
Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Thr Trp Ile
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Lys Ile Asn
His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val
Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys
Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90
95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met
100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala
Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr
Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr
Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro 210 215
220Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
Phe225 230 235 240Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val 245 250 255Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe 260 265 270Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr Lys Pro 275 280 285Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr 290 295 300Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val305 310 315 320Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 325 330
335Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
340 345 350Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
Lys Gly 355 360 365Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly Gln Pro 370 375 380Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp Ser Asp Gly Ser385 390 395 400Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly 435 440113442PRTArtificial
SequenceSynthetic 3C3 (VH + G2.g1.1) or 3C3-IgG2- IgG1.1 113Gln Val
Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr
Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25
30Tyr Trp Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile
35 40 45Gly Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu
Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val
Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp
Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys
Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser Thr
Ser Glu Ser Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala
Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170
175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn
180 185 190Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro
Ser Asn 195 200 205Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys
Val Glu Cys Pro 210 215 220Pro Cys Pro Ala Pro Pro Val Ala Gly Pro
Ser Val Phe Leu Phe Pro225 230 235 240Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr 245 250 255Cys Val Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 260 265 270Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 275 280 285Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 290 295
300Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser305 310 315 320Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile
Ser Lys Ala Lys 325 330 335Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
Leu Pro Pro Ser Arg Glu 340 345 350Glu Met Thr Lys Asn Gln Val Ser
Leu Thr Cys Leu Val Lys Gly Phe 355 360 365Tyr Pro Ser Asp Ile Ala
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 370 375 380Asn Asn Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe385 390 395 400Phe
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 405 410
415Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
420 425 430Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435
440114443PRTArtificial SequenceSynthetic 3C3 (VH + G2(C219S).g1) or
3C3-IgG2- C219S-IgG1 114Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu
Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly
Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Thr Trp Ile Arg Gln Pro Pro
Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Lys Ile Asn His Ser Gly Asn
Thr Asn Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val
Asp Thr Ser Lys Asn Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val
Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Leu Gly Ala
Phe Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105 110Val Thr
Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120
125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr Gln Thr Tyr Thr
Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr Lys Val Asp Lys
Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro 210 215 220Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe225 230 235
240Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
Lys Phe 260 265 270Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro 275 280 285Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
Val Val Ser Val Leu Thr 290 295 300Val Leu His Gln Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val305 310 315 320Ser Asn Lys Ala Leu
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 325 330 335Lys Gly Gln
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350Glu
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360
365Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
Gly Ser385 390 395 400Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
Ser Arg Trp Gln Gln 405 410 415Gly Asn Val Phe Ser Cys Ser Val Met
His Glu Ala Leu His Asn His 420 425 430Tyr Thr Gln Lys Ser Leu Ser
Leu Ser Pro Gly 435 440115442PRTArtificial SequenceSynthetic 3C3
(VH + G2(C219S).g1.1) or 3C3- IgG2-C219S-IgG1.1 115Gln Val Gln Leu
Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser
Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp
Thr Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly
Lys Ile Asn His Ser Gly Asn Thr Asn Tyr Asn Pro Ser Leu Lys 50 55
60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe Ser Leu65
70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
Ala 85 90 95Arg Leu Gly Ala Phe Asp Ala Phe Asp Ile Trp Gly Gln Gly
Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200
205Thr Lys Val Asp Lys Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro
210 215 220Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
Phe Pro225 230 235 240Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val Thr 245 250 255Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe Asn 260 265 270Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg 275 280 285Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 290 295 300Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser305 310 315
320Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys
325 330 335Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
Arg Glu 340 345 350Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
Val Lys Gly Phe 355 360 365Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu 370 375 380Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe385 390 395 400Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 405 410 415Asn Val Phe
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 420 425 430Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440116124PRTArtificial
SequenceSynthetic 2G6 (VH) 116Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp
Gly Ser Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115
120117107PRTArtificial
SequenceSynthetic 2G6 (VL) 117Asp Ile Gln Met Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 85 90 95Thr Phe
Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 105118450PRTArtificial
SequenceSynthetic 2G6 (full length wild-type heavy chain) 118Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Leu Ser Asp Tyr
20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Ser Asn Lys Phe Tyr Val Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Leu Ala Thr Gly His
Phe Tyr Tyr Val Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val
Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170
175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr
Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
Thr Val Glu Arg 210 215 220Lys Cys Cys Val Glu Cys Pro Pro Cys Pro
Ala Pro Pro Val Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg 290 295
300Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly Lys 450119214PRTArtificial SequenceSynthetic
2G6 (full length wild-type light chain) 119Asp Ile Gln Met Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile
Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr
Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala
Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr
85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala
Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala
Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr
Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 210120453PRTArtificial SequenceSynthetic
2G6.IgG1 (VH + IgG1) 120Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Ser
Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
130 135 140Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser
Leu Gly Thr Gln Thr Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu225 230 235
240Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
245 250 255Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp 260 265 270Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly 275 280 285Val Glu Val His Asn Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn 290 295 300Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val Leu His Gln Asp Trp305 310 315 320Leu Asn Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 325 330 335Ala Pro Ile
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360
365Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
370 375 380Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
Lys Thr385 390 395 400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys 405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys 420 425 430Ser Val Met His Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450121453PRTArtificial SequenceSynthetic 2G6.IgG1.1 (VH + IgG1.1)
121Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Leu Ser Asp
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Ser Asn Lys Phe Tyr Val
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Leu Ala Thr Gly
His Phe Tyr Tyr Val Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450122214PRTArtificial SequenceSynthetic 2G6.IgG1 (VL + CL) 122Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu
Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 2101235PRTArtificial
SequenceSynthetic 2G6 VH CDR1 123Asp Tyr Gly Met His1
512417PRTArtificial SequenceSynthetic 2G6 VH CDR2 124Val Ile Trp
Tyr Asp Gly Ser Asn Lys Phe Tyr Val Asp Ser Val Lys1 5 10
15Gly12515PRTArtificial SequenceSynthetic 2G6 VH CDR3 125Gly Gly
Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp Val1 5 10
1512611PRTArtificial SequenceSynthetic 2G6 VL CDR1 126Arg Ala Ser
Gln Gly Ile Ser Ser Trp Leu Ala1 5 101277PRTArtificial
SequenceSynthetic 2G6 VL CDR2 127Ala Ala Ser Ser Leu Gln Ser1
51289PRTArtificial SequenceSynthetic 2G6 VL CDR3 128Gln Gln Tyr Asn
Ser Tyr Pro Tyr Thr1 5129449PRTArtificial SequenceSynthetic 2G6 (VH
+ G2) or 2G6-IgG2 129Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Ser
Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Phe Asn Ser Thr Phe Arg 290 295 300Val Val Ser Val Leu Thr Val Val
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr Ile
Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Met385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe
Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly130449PRTArtificial
SequenceSynthetic 2G6 (VH + G2(C219S)) or2G6-IgG2- C219S 130Gln Val
Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Gly1 5 10 15Ser
Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile Leu Ser Asp Tyr 20 25
30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Thr Val Ile Trp Tyr Asp Gly Ser Asn Lys Phe Tyr Val Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Val Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Arg Leu Ala Thr Gly His Phe
Tyr Tyr Val Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr
Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu
Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170
175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr
Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
Thr Val Glu Arg 210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro
Ala Pro Pro Val Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg 290 295
300Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly131450PRTArtificial SequenceSynthetic 2G6 (VH
+ G2.g1) or2G6-IgG2-IgG1 131Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly
Ser Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235
240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360
365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450132449PRTArtificial SequenceSynthetic 2G6 (VH + G2.g1.1)
or2G6-IgG2- IgG1.1 132Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly Ser
Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly133450PRTArtificial SequenceSynthetic 2G6 (VH + G2(C219S).g1)
or 2G6-IgG2- C219S-IgG1 133Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly
Ser Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Ser Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235
240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360
365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450134449PRTArtificial SequenceSynthetic 2G6 (VH + G2(C219S).g1.1)
or2G6- IgG2-C219S-IgG1.1 134Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Ile Leu Ser Asp Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Thr Val Ile Trp Tyr Asp Gly
Ser Asn Lys Phe Tyr Val Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Arg Leu Ala Thr Gly His Phe Tyr Tyr Val Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Ser Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly135124PRTArtificial SequenceSynthetic 8A6 (VH) 135Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly
Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Leu Met Val Arg Gly Leu Phe Tyr
Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val
Ser Ser 115 120136107PRTArtificial SequenceSynthetic 8A6 (VL)
136Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val
Gly1
5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser
Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe
Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
Phe Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu
Ile Lys 100 105137450PRTArtificial SequenceSynthetic 8A6 (full
length wild-type heavy chain) 137Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr
Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met Gln Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr
Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230
235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Phe Asn Ser Thr Phe Arg 290 295 300Val Val Ser Val Leu Thr Val
Val His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr
Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Met385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445Gly Lys
450138214PRTArtificial SequenceSynthetic 8A6 (full length wild-type
light chain) 138Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly
Ile Ser Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala
Pro Lys Phe Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr
Cys Gln Gln Phe Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala
Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135
140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser
Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr
Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr
Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln Gly
Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys
210139453PRTArtificial SequenceSynthetic 8A6.IgG1 (VH + IgG1)
139Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser
Tyr 20 25 30Gly Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Leu Met Val Arg Gly
Leu Phe Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Leu Leu Gly Gly Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ala Pro Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450140453PRTArtificial SequenceSynthetic 8A6.IgG1.1 (VH + IgG1.1)
140Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser
Tyr 20 25 30Gly Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Leu Met Val Arg Gly
Leu Phe Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450141214PRTArtificial SequenceSynthetic 8A6.IgG1 (VL + CL) 141Ala
Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu
Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 2101425PRTArtificial
SequenceSynthetic 8A6 VH CDR1 142Ser Tyr Gly Met Gln1
514317PRTArtificial SequenceSynthetic 8A6 VH CDR2 143Val Ile Trp
Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val Lys1 5 10
15Gly14415PRTArtificial SequenceSynthetic 8A6 VH CDR3 144Gly Gly
Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp Val1 5 10
1514511PRTArtificial SequenceSynthetic 8A6 VL CDR1 145Arg Ala Ser
Gln Gly Ile Ser Ser Ala Leu Ala1 5 101467PRTArtificial
SequenceSynthetic 8A6 VL CDR2 146Asp Ala Ser Ser Leu Glu Ser1
51479PRTArtificial SequenceSynthetic 8A6 VL CDR3 147Gln Gln Phe Asn
Ser Tyr Pro Tyr Thr1 5148449PRTArtificial SequenceSynthetic 8A6 (VH
+ G2(C219S)) or 8A6-IgG2- C219S 148Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr
Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met Gln Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr
Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230
235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Phe Asn Ser Thr Phe Arg 290 295 300Val Val Ser Val Leu Thr Val
Val His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile Glu 325 330 335Lys Thr
Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Met385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser
Leu Ser Leu Ser Pro 435 440 445Gly149450PRTArtificial
SequenceSynthetic 8A6 (VH + G2.g1) or 8A6 -IgG2-IgG1 149Gln Val Gln
Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu
Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly
Met Gln Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Ala Val Ile Trp Tyr Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val
50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu
Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val
Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Leu Met Val Arg Gly Leu Phe Tyr
Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val
Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala
Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly
Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val
Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe
Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185
190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn
195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val
Glu Arg 210 215 220Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro
Glu Leu Leu Gly225 230 235 240Gly Pro Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr Leu Met 245 250 255Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val Val Asp Val Ser His 260 265 270Glu Asp Pro Glu Val
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg
Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310
315 320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
Ile 325 330 335Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu
Pro Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
Lys Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425
430His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445Pro Gly 450150449PRTArtificial SequenceSynthetic 8A6 (VH
+ G2.g1.1) or 8A6 -IgG2- IgG1.1 150Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr
Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met Gln Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr
Glu Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230
235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly151450PRTArtificial SequenceSynthetic 8A6 (VH + G2(C219S).g1)
or 8A6-IgG2- C219S-IgG1 151Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr Ala Ser
Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met Gln Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu Gly
Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Glu Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Ser Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230 235
240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360
365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu
370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr
Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450152449PRTArtificial SequenceSynthetic 8A6 (VH + G2(C219S).g1.1)
or 8A6- IgG2-C219S-IgG1.1 152Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Thr Ala
Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30Gly Met Gln Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Glu
Gly Ser Asn Lys Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Glu Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg
Gly Gly Leu Met Val Arg Gly Leu Phe Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230
235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met
Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val
Ser His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val
Leu His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly153125PRTArtificial SequenceSynthetic 9G7 (VH) 153Glu Val Gln
Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu
Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Val 20 25 30Trp
Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala
50 55 60Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn
Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu His Thr Glu Asp Thr
Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly Gln Leu Ile Pro Tyr Ser Tyr
Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Ser Val Thr
Val Ser Ser 115 120 125154108PRTArtificial SequenceSynthetic 9G7
(VL1) 154Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser
Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val
Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala
Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile
Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr
Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys 100 105155108PRTArtificial SequenceSynthetic
9G7 (VL2) 155Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu
Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
Val Thr Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly
Ile Pro Glu Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr
Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Ile Thr Phe Gly Gln Gly
Thr Arg Leu Glu Ile Lys 100 105156452PRTArtificial
SequenceSynthetic 9G7 (full length wild-type heavy chain) 156Glu
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Val
20 25 30Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr
Ala Ala 50 55 60Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser
Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu His Thr Glu
Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly Gln Leu Ile Pro Tyr
Ser Tyr Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Ser
Val Thr Val Ser Ser Ala Ser Thr 115 120 125Lys Gly Pro Ser Val Phe
Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140Glu Ser Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu145 150 155 160Pro
Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170
175Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
180 185 190Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Lys Thr Tyr
Thr Cys 195 200 205Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp
Lys Arg Val Glu 210 215 220Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys
Pro Ala Pro Glu Phe Leu225 230 235 240Gly Gly Pro Ser
Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255Met Ile
Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265
270Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu
275 280 285Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn
Ser Thr 290 295 300Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn305 310 315 320Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Gly Leu Pro Ser Ser 325 330 335Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350Val Tyr Thr Leu Pro
Pro Ser Gln Glu Glu Met Thr Lys Asn Gln Val 355 360 365Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro385 390
395 400Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu
Thr 405 410 415Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser
Cys Ser Val 420 425 430Met His Glu Ala Leu His Asn His Tyr Thr Gln
Lys Ser Leu Ser Leu 435 440 445Ser Leu Gly Lys
450157215PRTArtificial SequenceSynthetic 9G7 L2 (full length
wild-type light chain 2) 157Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala
Ser Gln Ser Val Thr Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg
Ala Thr Gly Ile Pro Glu Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe
Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Ile Thr Phe
Gly Gln Gly Thr Arg Leu Glu Ile Lys Arg Thr Val Ala 100 105 110Ala
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120
125Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
Asn Ser145 150 155 160Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
Ser Thr Tyr Ser Leu 165 170 175Ser Ser Thr Leu Thr Leu Ser Lys Ala
Asp Tyr Glu Lys His Lys Val 180 185 190Tyr Ala Cys Glu Val Thr His
Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205Ser Phe Asn Arg Gly
Glu Cys 210 215158454PRTArtificial SequenceSynthetic 9G7.IgG1 (VH +
IgG1) 158Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro
Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe
Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly
Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr
Thr Asp Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg Phe Thr Ile Ser Arg
Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu
His Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly Gln Leu
Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln
Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr 115 120 125Lys Gly Pro
Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 130 135 140Gly
Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu145 150
155 160Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val
His 165 170 175Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser
Leu Ser Ser 180 185 190Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
Gln Thr Tyr Ile Cys 195 200 205Asn Val Asn His Lys Pro Ser Asn Thr
Lys Val Asp Lys Arg Val Glu 210 215 220Pro Lys Ser Cys Asp Lys Thr
His Thr Cys Pro Pro Cys Pro Ala Pro225 230 235 240Glu Leu Leu Gly
Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 245 250 255Asp Thr
Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 260 265
270Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
275 280 285Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu
Gln Tyr 290 295 300Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val
Leu His Gln Asp305 310 315 320Trp Leu Asn Gly Lys Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu 325 330 335Pro Ala Pro Ile Glu Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg 340 345 350Glu Pro Gln Val Tyr
Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 355 360 365Asn Gln Val
Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 370 375 380Ile
Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys385 390
395 400Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr
Ser 405 410 415Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn
Val Phe Ser 420 425 430Cys Ser Val Met His Glu Ala Leu His Asn His
Tyr Thr Gln Lys Ser 435 440 445Leu Ser Leu Ser Pro Gly
450159454PRTArtificial SequenceSynthetic 9G7.IgG1.1 (VH + IgG1.1)
159Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr
Val 20 25 30Trp Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp
Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp
Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu His Thr
Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly Gln Leu Ile Pro
Tyr Ser Tyr Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr
Ser Val Thr Val Ser Ser Ala Ser Thr 115 120 125Lys Gly Pro Ser Val
Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 130 135 140Gly Gly Thr
Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu145 150 155
160Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
165 170 175Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu
Ser Ser 180 185 190Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln
Thr Tyr Ile Cys 195 200 205Asn Val Asn His Lys Pro Ser Asn Thr Lys
Val Asp Lys Arg Val Glu 210 215 220Pro Lys Ser Cys Asp Lys Thr His
Thr Cys Pro Pro Cys Pro Ala Pro225 230 235 240Glu Ala Glu Gly Ala
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 245 250 255Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 260 265 270Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 275 280
285Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
290 295 300Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His
Gln Asp305 310 315 320Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser Asn Lys Ala Leu 325 330 335Pro Ser Ser Ile Glu Lys Thr Ile Ser
Lys Ala Lys Gly Gln Pro Arg 340 345 350Glu Pro Gln Val Tyr Thr Leu
Pro Pro Ser Arg Glu Glu Met Thr Lys 355 360 365Asn Gln Val Ser Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 370 375 380Ile Ala Val
Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys385 390 395
400Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
405 410 415Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser 420 425 430Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser 435 440 445Leu Ser Leu Ser Pro Gly
450160215PRTArtificial SequenceSynthetic 9G7.IgG1 (VL1 + CL) 160Glu
Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10
15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg
Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe Gly Gln Gly Thr Lys Val Glu
Ile Lys Arg Thr Val Ala 100 105 110Ala Pro Ser Val Phe Ile Phe Pro
Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125Gly Thr Ala Ser Val Val
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140Ala Lys Val Gln
Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser145 150 155 160Gln
Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170
175Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val
180 185 190Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val
Thr Lys 195 200 205Ser Phe Asn Arg Gly Glu Cys 210
215161215PRTArtificial SequenceSynthetic 9G7.IgG1.2 (VL2 + CL)
161Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1
5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Thr Ser
Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Glu
Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln
Gln Tyr Gly Ser Ser Pro 85 90 95Ile Thr Phe Gly Gln Gly Thr Arg Leu
Glu Ile Lys Arg Thr Val Ala 100 105 110Ala Pro Ser Val Phe Ile Phe
Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125Gly Thr Ala Ser Val
Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140Ala Lys Val
Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser145 150 155
160Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu
165 170 175Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His
Lys Val 180 185 190Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
Pro Val Thr Lys 195 200 205Ser Phe Asn Arg Gly Glu Cys 210
2151625PRTArtificial SequenceSynthetic 9G7 VH CDR1 162Thr Val Trp
Met Ser1 516319PRTArtificial SequenceSynthetic 9G7 VH CDR2 163Arg
Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala Pro1 5 10
15Val Lys Gly16414PRTArtificial SequenceSynthetic 9G7 VH CDR3
164Gly Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met Asp Val1 5
1016512PRTArtificial SequenceSynthetic 9G7 VL1 CDR1 165Arg Ala Ser
Gln Ser Val Ser Ser Ser Tyr Leu Ala1 5 101667PRTArtificial
SequenceSynthetic 9G7 VL1 CDR2 166Gly Ala Ser Ser Arg Ala Thr1
51679PRTArtificial SequenceSynthetic 9G7 VL1 CDR3 167Gln Gln Tyr
Gly Ser Ser Pro Trp Thr1 516812PRTArtificial SequenceSynthetic 9G7
VL2 CDR1 168Arg Ala Ser Gln Ser Val Thr Ser Ser Tyr Leu Ala1 5
101697PRTArtificial SequenceSynthetic 9G7 VL2 CDR2 169Gly Ala Ser
Ser Arg Ala Thr1 51709PRTArtificial SequenceSynthetic 9G7 VL2 CDR3
170Gln Gln Tyr Gly Ser Ser Pro Ile Thr1 5171450PRTArtificial
SequenceSynthetic 9G7 (VH + G2) or 9G7-IgG2 171Glu Val Gln Leu Val
Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Val 20 25 30Trp Met Ser
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg
Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60Pro
Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75
80Leu Tyr Leu Gln Met Asn Ser Leu His Thr Glu Asp Thr Ala Val Tyr
85 90 95Tyr Cys Thr Thr Gly Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly
Met 100 105 110Asp Val Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser
Ala Ser Thr 115 120 125Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys
Ser Arg Ser Thr Ser 130 135 140Glu Ser Thr Ala Ala Leu Gly Cys Leu
Val Lys Asp Tyr Phe Pro Glu145 150 155 160Pro Val Thr Val Ser Trp
Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175Thr Phe Pro Ala
Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190Val Val
Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys 195 200
205Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu
210 215 220Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro
Val Ala225 230 235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His 260 265 270Glu Asp Pro Glu Val Gln Phe
Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr
Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe 290 295 300Arg Val Val
Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly305 310 315
320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro Ile
325 330 335Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro
Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400Met Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440
445Pro Gly 450172450PRTArtificial SequenceSynthetic 9G7 (VH +
G2(C219S)) or 9G7-IgG2- C219S 172Glu Val Gln Leu Val Glu Ser Gly
Gly Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40
45Gly Arg Ile Lys Ser Lys Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala
50 55 60Pro Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn
Thr65 70 75 80Leu Tyr Leu Gln Met Asn Ser Leu His Thr Glu Asp Thr
Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly Gln Leu Ile Pro Tyr Ser Tyr
Tyr Tyr Gly Met 100 105 110Asp Val Trp Gly Gln Gly Thr Ser Val Thr
Val Ser Ser Ala Ser Thr 115 120 125Lys Gly Pro Ser Val Phe Pro Leu
Ala Pro Cys Ser Arg Ser Thr Ser 130 135 140Glu Ser Thr Ala Ala Leu
Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu145 150 155 160Pro Val Thr
Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 165 170 175Thr
Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185
190Val Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys
195 200 205Asn Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr
Val Glu 210 215 220Arg Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala
Pro Pro Val Ala225 230 235 240Gly Pro Ser Val Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr Leu Met 245 250 255Ile Ser Arg Thr Pro Glu Val
Thr Cys Val Val Val Asp Val Ser His 260 265 270Glu Asp Pro Glu Val
Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285His Asn Ala
Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe 290 295 300Arg
Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly305 310
315 320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro
Ile 325 330 335Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu
Pro Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr
Lys Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400Met Leu Asp Ser
Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys
Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425
430His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
435 440 445Pro Gly 450173451PRTArtificial SequenceSynthetic 9G7 (VH
+ G2.g1) or 9G7-IgG2-IgG1 173Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys
Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu
Gln Met Asn Ser Leu His Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys
Thr Thr Gly Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met 100 105
110Asp Val Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr
115 120 125Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser 130 135 140Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu145 150 155 160Pro Val Thr Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His 165 170 175Thr Phe Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190Val Val Thr Val Pro
Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys 195 200 205Asn Val Asp
His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu 210 215 220Arg
Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu225 230
235 240Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr
Leu 245 250 255Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
Asp Val Ser 260 265 270His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr
Val Asp Gly Val Glu 275 280 285Val His Asn Ala Lys Thr Lys Pro Arg
Glu Glu Gln Tyr Asn Ser Thr 290 295 300Tyr Arg Val Val Ser Val Leu
Thr Val Leu His Gln Asp Trp Leu Asn305 310 315 320Gly Lys Glu Tyr
Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335Ile Glu
Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345
350Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val
355 360 365Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
Ala Val 370 375 380Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
Lys Thr Thr Pro385 390 395 400Pro Val Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys Leu Thr 405 410 415Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430Met His Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445Ser Pro Gly
450174450PRTArtificial SequenceSynthetic 9G7 (VH + G2.g1.1) or
9G7-IgG2- IgG1.1 174Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val
Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe
Thr Phe Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg Gln Ala Pro Gly
Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Thr Asp Gly
Gly Thr Thr Asp Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met Asn
Ser Leu His Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Thr Gly
Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met 100 105 110Asp Val Trp
Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr 115 120 125Lys
Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser 130 135
140Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu145 150 155 160Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His 165 170 175Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser 180 185 190Val Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys 195 200 205Asn Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu 210 215 220Arg Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala225 230 235 240Gly
Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 245 250
255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His
260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val
Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr Val Leu His
Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys Cys Lys Val
Ser Asn Lys Ala Leu Pro Ser Ser Ile 325 330 335Glu Lys Thr Ile Ser
Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345 350Tyr Thr Leu
Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser 355 360 365Leu
Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 370 375
380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450175451PRTArtificial SequenceSynthetic 9G7 (VH + G2(C219S).g1) or
9G7-IgG2- C219S-IgG1 175Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu
Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys Thr Asp
Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu Gln Met
Asn Ser Leu His Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys Thr Thr
Gly Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met 100 105 110Asp Val
Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr 115 120
125Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser
130 135 140Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe
Pro Glu145 150 155 160Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
Thr Ser Gly Val His 165 170 175Thr Phe Pro Ala Val Leu Gln Ser Ser
Gly Leu Tyr Ser Leu Ser Ser 180 185 190Val Val Thr Val Pro Ser Ser
Asn Phe Gly Thr Gln Thr Tyr Thr Cys 195 200 205Asn Val Asp His Lys
Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu 210 215 220Arg Lys Ser
Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu225 230 235
240Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
245 250 255Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser 260 265 270His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu 275 280 285Val His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr 290 295 300Tyr Arg Val Val Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn305 310 315 320Gly Lys Glu Tyr Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 325 330 335Ile Glu Lys
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350Val
Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 355 360
365Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
370 375 380Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro385 390 395 400Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr 405 410 415Val Asp Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val 420 425 430Met His Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445Ser Pro Gly
450176450PRTArtificial SequenceSynthetic 9G7 (VH + G2(C219S).g1.1)
or 9G7- IgG2-C219S-IgG1.1 176Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Lys Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Thr Val 20 25 30Trp Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Gly Arg Ile Lys Ser Lys
Thr Asp Gly Gly Thr Thr Asp Tyr Ala Ala 50 55 60Pro Val Lys Gly Arg
Phe Thr Ile Ser Arg Asp Asp Ser Lys Asn Thr65 70 75 80Leu Tyr Leu
Gln Met Asn Ser Leu His Thr Glu Asp Thr Ala Val Tyr 85 90 95Tyr Cys
Thr Thr Gly Gln Leu Ile Pro Tyr Ser Tyr Tyr Tyr Gly Met 100 105
110Asp Val Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser Thr
115 120 125Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser
Thr Ser 130 135 140Glu Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp
Tyr Phe Pro Glu145 150 155 160Pro Val Thr Val Ser Trp Asn Ser Gly
Ala Leu Thr Ser Gly Val His 165 170 175Thr Phe Pro Ala Val Leu Gln
Ser Ser Gly Leu Tyr Ser Leu Ser Ser 180 185 190Val Val Thr Val Pro
Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys 195 200 205Asn Val Asp
His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu 210 215 220Arg
Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala225 230
235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile 325 330 335Glu Lys
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450177215PRTArtificial SequenceSynthetic 9G7 L1 (full length
wild-type light chain 1) 177Glu Ile Val Leu Thr Gln Ser Pro Gly Thr
Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala
Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg
Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe
Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Trp Thr Phe
Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110Ala
Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120
125Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu
130 135 140Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly
Asn Ser145 150 155 160Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp
Ser Thr Tyr Ser Leu 165 170 175Ser Ser Thr Leu Thr Leu Ser Lys Ala
Asp Tyr Glu Lys His Lys Val 180 185 190Tyr Ala Cys Glu Val Thr His
Gln Gly Leu Ser Ser Pro Val Thr Lys 195
200 205Ser Phe Asn Arg Gly Glu Cys 210 215178117PRTArtificial
SequenceSynthetic 14E3 (VH) 178Gln Val Gln Leu Gln Gln Trp Gly Ala
Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val
Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln
Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Ser
Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile
Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys Leu Ser
Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Phe
Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105
110Val Thr Val Ser Ser 115179107PRTArtificial SequenceSynthetic
14E3 (VL) 179Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly
Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala
Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr
Cys Gln Gln Tyr Asn Ser Tyr Pro Pro 85 90 95Thr Phe Gly Gln Gly Thr
Lys Val Glu Ile Lys 100 105180447PRTArtificial SequenceSynthetic
14E3 (full length wild-type heavy chain) 180Gln Val Gln Leu Gln Gln
Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr
Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp
Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile
Asn His Ser Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg
Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75
80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95Arg Phe Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr
Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
Phe Pro Leu 115 120 125Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr
Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro
Val Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly
Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190Leu Gly
Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200
205Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His
210 215 220Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro
Ser Val225 230 235 240Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr 245 250 255Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His Glu Asp Pro Glu 260 265 270Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala Lys 275 280 285Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser 290 295 300Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys305 310 315
320Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile
325 330 335Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
Leu Pro 340 345 350Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser
Leu Thr Cys Leu 355 360 365Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu Trp Glu Ser Asn 370 375 380Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser385 390 395 400Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg 405 410 415Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu 420 425 430His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440
445181214PRTArtificial SequenceSynthetic 14E3 (full length
wild-type light chain) 181Asp Ile Gln Met Thr Gln Ser Pro Ser Ser
Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala
Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro
Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala Ser Ser Leu Gln
Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala
Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Pro 85 90 95Thr Phe Gly
Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110Pro
Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120
125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala
130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn
Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp
Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln
Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu
Cys 210182446PRTArtificial SequenceSynthetic 14E3.IgG1 (VH + IgG1)
182Gln Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1
5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly
Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
Trp Ile 35 40 45Gly Glu Ile Asn His Ser Gly Asn Thr Tyr Tyr Asn Pro
Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn
Gln Leu Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr
Ala Val Tyr Tyr Cys Ala 85 90 95Arg Phe Gly Ser Asn Asp Ala Phe Asp
Ile Trp Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser
Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Ser Ser Lys
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys
Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155
160Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser
165 170 175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser
Ser Ser 180 185 190Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His
Lys Pro Ser Asn 195 200 205Thr Lys Val Asp Lys Arg Val Glu Pro Lys
Ser Cys Asp Lys Thr His 210 215 220Thr Cys Pro Pro Cys Pro Ala Pro
Glu Leu Leu Gly Gly Pro Ser Val225 230 235 240Phe Leu Phe Pro Pro
Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr 245 250 255Pro Glu Val
Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu 260 265 270Val
Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys 275 280
285Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser
290 295 300Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu
Tyr Lys305 310 315 320Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro
Ile Glu Lys Thr Ile 325 330 335Ser Lys Ala Lys Gly Gln Pro Arg Glu
Pro Gln Val Tyr Thr Leu Pro 340 345 350Pro Ser Arg Glu Glu Met Thr
Lys Asn Gln Val Ser Leu Thr Cys Leu 355 360 365Val Lys Gly Phe Tyr
Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn 370 375 380Gly Gln Pro
Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser385 390 395
400Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg
405 410 415Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu
Ala Leu 420 425 430His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser
Pro Gly 435 440 445183446PRTArtificial SequenceSynthetic
14E3.IgG1.1 (VH + IgG1.1) 183Gln Val Gln Leu Gln Gln Trp Gly Ala
Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val
Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln
Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Ser
Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile
Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys Leu Ser
Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Phe
Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105
110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu
Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser Ser 180 185 190Leu Gly Thr Gln Thr
Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn 195 200 205Thr Lys Val
Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys Thr His 210 215 220Thr
Cys Pro Pro Cys Pro Ala Pro Glu Ala Glu Gly Ala Pro Ser Val225 230
235 240Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr 245 250 255Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu 260 265 270Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys 275 280 285Thr Lys Pro Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser 290 295 300Val Leu Thr Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys305 310 315 320Cys Lys Val Ser
Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile 325 330 335Ser Lys
Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro 340 345
350Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
355 360 365Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser Asn 370 375 380Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser385 390 395 400Asp Gly Ser Phe Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys Ser Arg 405 410 415Trp Gln Gln Gly Asn Val Phe
Ser Cys Ser Val Met His Glu Ala Leu 420 425 430His Asn His Tyr Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445184214PRTArtificial
SequenceSynthetic 14E3.IgG1 (VL + CL) 184Asp Ile Gln Met Thr Gln
Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile
Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Trp 20 25 30Leu Ala Trp Tyr
Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 35 40 45Tyr Ala Ala
Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly
Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Pro
85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala
Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu
Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn Ala
Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu Gln
Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu Thr
Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala Cys
Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 2101855PRTArtificial SequenceSynthetic
14E3 VH CDR1 185Gly Tyr Tyr Trp Ser1 518616PRTArtificial
SequenceSynthetic 14E3 VH CDR2 186Glu Ile Asn His Ser Gly Asn Thr
Tyr Tyr Asn Pro Ser Leu Lys Ser1 5 10 151879PRTArtificial
SequenceSynthetic 14E3 VH CDR3 187Phe Gly Ser Asn Asp Ala Phe Asp
Ile1 518811PRTArtificial SequenceSynthetic 14E3 VL CDR1 188Arg Ala
Ser Gln Gly Ile Ser Ser Trp Leu Ala1 5 101897PRTArtificial
SequenceSynthetic 14E3 VL CDR2 189Ala Ala Ser Ser Leu Gln Ser1
51909PRTArtificial SequenceSynthetic 14E3 VL CDR3 190Gln Gln Tyr
Asn Ser Tyr Pro Pro Thr1 5191442PRTArtificial SequenceSynthetic
14E3 (VH + G2) or 14E3-IgG2 191Gln Val Gln Leu Gln Gln Trp Gly Ala
Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val
Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln
Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Ser
Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile
Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys Leu Ser
Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Phe
Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105
110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr Gln Thr
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr Lys Val
Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro 210 215 220Pro
Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro225 230
235 240Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr 245 250 255Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
Gln Phe Asn 260 265 270Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro Arg 275 280 285Glu Glu Gln Phe Asn Ser Thr Phe Arg
Val Val Ser Val Leu Thr Val 290 295 300Val His Gln Asp Trp Leu Asn
Gly
Lys Glu Tyr Lys Cys Lys Val Ser305 310 315 320Asn Lys Gly Leu Pro
Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys 325 330 335Gly Gln Pro
Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 340 345 350Glu
Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe 355 360
365Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu
370 375 380Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser Asp Gly
Ser Phe385 390 395 400Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser
Arg Trp Gln Gln Gly 405 410 415Asn Val Phe Ser Cys Ser Val Met His
Glu Ala Leu His Asn His Tyr 420 425 430Thr Gln Lys Ser Leu Ser Leu
Ser Pro Gly 435 440192442PRTArtificial SequenceSynthetic 14E3 (VH +
G2(C219S)) or 14E3-IgG2- C219S 192Gln Val Gln Leu Gln Gln Trp Gly
Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala
Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg
Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His
Ser Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr
Ile Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys Leu
Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg
Phe Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105
110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu
115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu
Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val
Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr
Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser
Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr Gln Thr
Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr Lys Val
Asp Lys Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro 210 215 220Pro
Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro225 230
235 240Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
Thr 245 250 255Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
Gln Phe Asn 260 265 270Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro Arg 275 280 285Glu Glu Gln Phe Asn Ser Thr Phe Arg
Val Val Ser Val Leu Thr Val 290 295 300Val His Gln Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val Ser305 310 315 320Asn Lys Gly Leu
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Thr Lys 325 330 335Gly Gln
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu 340 345
350Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe
355 360 365Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
Pro Glu 370 375 380Asn Asn Tyr Lys Thr Thr Pro Pro Met Leu Asp Ser
Asp Gly Ser Phe385 390 395 400Phe Leu Tyr Ser Lys Leu Thr Val Asp
Lys Ser Arg Trp Gln Gln Gly 405 410 415Asn Val Phe Ser Cys Ser Val
Met His Glu Ala Leu His Asn His Tyr 420 425 430Thr Gln Lys Ser Leu
Ser Leu Ser Pro Gly 435 440193443PRTArtificial SequenceSynthetic
14E3 (VH + G2.g1) or 14E3-IgG2-IgG1 193Gln Val Gln Leu Gln Gln Trp
Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys
Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile
Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn
His Ser Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val
Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys
Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90
95Arg Phe Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met
100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe
Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala
Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val
Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val
His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser
Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr
Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr
Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys Val Glu Cys Pro 210 215
220Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
Phe225 230 235 240Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val 245 250 255Thr Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe 260 265 270Asn Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr Lys Pro 275 280 285Arg Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr 290 295 300Val Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val305 310 315 320Ser
Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 325 330
335Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
340 345 350Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val
Lys Gly 355 360 365Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser
Asn Gly Gln Pro 370 375 380Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
Leu Asp Ser Asp Gly Ser385 390 395 400Phe Phe Leu Tyr Ser Lys Leu
Thr Val Asp Lys Ser Arg Trp Gln Gln 405 410 415Gly Asn Val Phe Ser
Cys Ser Val Met His Glu Ala Leu His Asn His 420 425 430Tyr Thr Gln
Lys Ser Leu Ser Leu Ser Pro Gly 435 440194442PRTArtificial
SequenceSynthetic 14E3 (VH + G2.g1.1) or 14E3-IgG2- IgG1.1 194Gln
Val Gln Leu Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10
15Thr Leu Ser Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr
20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
Ile 35 40 45Gly Glu Ile Asn His Ser Gly Asn Thr Tyr Tyr Asn Pro Ser
Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln
Leu Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Phe Gly Ser Asn Asp Ala Phe Asp Ile
Trp Gly Gln Gly Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr
Lys Gly Pro Ser Val Phe Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser
Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly
Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170
175Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn
180 185 190Phe Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro
Ser Asn 195 200 205Thr Lys Val Asp Lys Thr Val Glu Arg Lys Cys Cys
Val Glu Cys Pro 210 215 220Pro Cys Pro Ala Pro Pro Val Ala Gly Pro
Ser Val Phe Leu Phe Pro225 230 235 240Pro Lys Pro Lys Asp Thr Leu
Met Ile Ser Arg Thr Pro Glu Val Thr 245 250 255Cys Val Val Val Asp
Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 260 265 270Trp Tyr Val
Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 275 280 285Glu
Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 290 295
300Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
Ser305 310 315 320Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile
Ser Lys Ala Lys 325 330 335Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr
Leu Pro Pro Ser Arg Glu 340 345 350Glu Met Thr Lys Asn Gln Val Ser
Leu Thr Cys Leu Val Lys Gly Phe 355 360 365Tyr Pro Ser Asp Ile Ala
Val Glu Trp Glu Ser Asn Gly Gln Pro Glu 370 375 380Asn Asn Tyr Lys
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe385 390 395 400Phe
Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 405 410
415Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr
420 425 430Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435
440195443PRTArtificial SequenceSynthetic 14E3 (VH + G2(C219S).g1)
or 14E3- IgG2-C219S-IgG1 195Gln Val Gln Leu Gln Gln Trp Gly Ala Gly
Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Ala Val Tyr
Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro
Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly Glu Ile Asn His Ser Gly
Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser
Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65 70 75 80Lys Leu Ser Ser
Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Phe Gly
Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly Thr Met 100 105 110Val
Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu 115 120
125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu Gly Cys
130 135 140Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp
Asn Ser145 150 155 160Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro
Ala Val Leu Gln Ser 165 170 175Ser Gly Leu Tyr Ser Leu Ser Ser Val
Val Thr Val Pro Ser Ser Asn 180 185 190Phe Gly Thr Gln Thr Tyr Thr
Cys Asn Val Asp His Lys Pro Ser Asn 195 200 205Thr Lys Val Asp Lys
Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro 210 215 220Pro Cys Pro
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe225 230 235
240Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
245 250 255Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val
Lys Phe 260 265 270Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala
Lys Thr Lys Pro 275 280 285Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
Val Val Ser Val Leu Thr 290 295 300Val Leu His Gln Asp Trp Leu Asn
Gly Lys Glu Tyr Lys Cys Lys Val305 310 315 320Ser Asn Lys Ala Leu
Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 325 330 335Lys Gly Gln
Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 340 345 350Glu
Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 355 360
365Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
370 375 380Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp
Gly Ser385 390 395 400Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys
Ser Arg Trp Gln Gln 405 410 415Gly Asn Val Phe Ser Cys Ser Val Met
His Glu Ala Leu His Asn His 420 425 430Tyr Thr Gln Lys Ser Leu Ser
Leu Ser Pro Gly 435 440196442PRTArtificial SequenceSynthetic 14E3
(VH + G2(C219S).g1.1) or 14E3- IgG2-C219S-IgG1.1 196Gln Val Gln Leu
Gln Gln Trp Gly Ala Gly Leu Leu Lys Pro Ser Glu1 5 10 15Thr Leu Ser
Leu Thr Cys Ala Val Tyr Gly Gly Ser Phe Ser Gly Tyr 20 25 30Tyr Trp
Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp Ile 35 40 45Gly
Glu Ile Asn His Ser Gly Asn Thr Tyr Tyr Asn Pro Ser Leu Lys 50 55
60Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Leu Ser Leu65
70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys
Ala 85 90 95Arg Phe Gly Ser Asn Asp Ala Phe Asp Ile Trp Gly Gln Gly
Thr Met 100 105 110Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser
Val Phe Pro Leu 115 120 125Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser
Thr Ala Ala Leu Gly Cys 130 135 140Leu Val Lys Asp Tyr Phe Pro Glu
Pro Val Thr Val Ser Trp Asn Ser145 150 155 160Gly Ala Leu Thr Ser
Gly Val His Thr Phe Pro Ala Val Leu Gln Ser 165 170 175Ser Gly Leu
Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Asn 180 185 190Phe
Gly Thr Gln Thr Tyr Thr Cys Asn Val Asp His Lys Pro Ser Asn 195 200
205Thr Lys Val Asp Lys Thr Val Glu Arg Lys Ser Cys Val Glu Cys Pro
210 215 220Pro Cys Pro Ala Pro Pro Val Ala Gly Pro Ser Val Phe Leu
Phe Pro225 230 235 240Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg
Thr Pro Glu Val Thr 245 250 255Cys Val Val Val Asp Val Ser His Glu
Asp Pro Glu Val Lys Phe Asn 260 265 270Trp Tyr Val Asp Gly Val Glu
Val His Asn Ala Lys Thr Lys Pro Arg 275 280 285Glu Glu Gln Tyr Asn
Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 290 295 300Leu His Gln
Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser305 310 315
320Asn Lys Ala Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys
325 330 335Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
Arg Glu 340 345 350Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu
Val Lys Gly Phe 355 360 365Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu
Ser Asn Gly Gln Pro Glu 370 375 380Asn Asn Tyr Lys Thr Thr Pro Pro
Val Leu Asp Ser Asp Gly Ser Phe385 390 395 400Phe Leu Tyr Ser Lys
Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly 405 410 415Asn Val Phe
Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 420 425 430Thr
Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440197124PRTArtificial
SequenceSynthetic 19H8 (VH) 197Gln Val Gln Leu Val Glu Ser Gly Gly
Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val Ile Trp Tyr Gly
Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn
Ser Lys Asn Ser Leu Ser65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ala Met Val
Arg Gly Val Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly
Thr Thr Val Thr Val Ser Ser 115 120198107PRTArtificial
SequenceSynthetic 19H8 (VL1) 198Ala Ile Gln Leu Thr Gln Ser Pro Ser
Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Phe Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu
Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro Gln 85 90 95Thr Phe
Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105199107PRTArtificial
SequenceSynthetic 19H8 (VL2) 199Glu Ile Val Leu Thr Gln Ser Pro Ala
Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg
Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg
Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75 80Glu Asp Phe
Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Leu 85 90 95Thr Phe
Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105200450PRTArtificial
SequenceSynthetic 19H8 (full length wild-type heavy chain) 200Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Met 35 40 45Ala Val Ile Trp Tyr Gly Gly Ser Asn Lys Phe Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Ser Leu Ser65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ala Met Val Arg Gly Val
Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val
Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170
175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr
Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
Thr Val Glu Arg 210 215 220Lys Cys Cys Val Glu Cys Pro Pro Cys Pro
Ala Pro Pro Val Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg 290 295
300Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly Lys 450201214PRTArtificial SequenceSynthetic
19H8 L1 (full length wild-type light chain 1) 201Ala Ile Gln Leu
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile 35 40 45Tyr
Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Ser Tyr Pro
Gln 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val
Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn
Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn
Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 210202214PRTArtificial SequenceSynthetic
19H8 L2 (full length wild-type light chain 2) 202Glu Ile Val Leu
Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr
Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65
70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro
Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val
Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln
Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys Leu Leu Asn Asn
Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp Asn
Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr Glu
Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr Leu
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190Ala
Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200
205Phe Asn Arg Gly Glu Cys 210203453PRTArtificial SequenceSynthetic
19H8.IgG1 (VH + IgG1) 203Gln Val Gln Leu Val Glu Ser Gly Gly Gly
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val Ile Trp Tyr Gly Gly
Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Ser Leu Ser65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
130 135 140Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser
Leu Gly Thr Gln Thr Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu225 230 235
240Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
245 250 255Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp 260 265 270Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly 275 280 285Val Glu Val His Asn Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn 290 295 300Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val Leu His Gln Asp Trp305 310 315 320Leu Asn Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 325 330 335Ala Pro Ile
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360
365Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
370 375 380Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
Lys Thr385 390 395 400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys 405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys 420 425 430Ser Val Met His Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450204453PRTArtificial SequenceSynthetic 19H8.IgG1.1 (VH + IgG1.1)
204Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Met 35 40 45Ala Val Ile Trp Tyr Gly Gly Ser Asn Lys Phe Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Ser Leu Ser65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ala Met Val Arg Gly
Val Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450205214PRTArtificial SequenceSynthetic 19H8.IgG1 (VL1 + CL)
205Ala Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1
5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser
Ala 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe
Leu Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln
Phe Asn Ser Tyr Pro Gln 85 90 95Thr Phe Gly Gln Gly Thr Lys Val Glu
Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro
Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val
Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln
Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155
160Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser
165 170 175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys
Val Tyr 180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro
Val Thr Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys
210206214PRTArtificial SequenceSynthetic 19H8.IgG1.2 (VL2 + CL)
206Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1
5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser
Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu
Leu Ile 35 40 45Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg
Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
Ser Leu Glu Pro65 70 75 80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln
Arg Ser Asn Trp Pro Leu 85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu
Ile Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro
Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val
Cys Leu Leu Asn
Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp Lys Val Asp
Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu Ser Val Thr
Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175Ser Thr
Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185
190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser
195 200 205Phe Asn Arg Gly Glu Cys 2102075PRTArtificial
SequenceSynthetic 19H8 VH CDR1 207Asn Tyr Gly Met His1
520817PRTArtificial SequenceSynthetic 19H8 VH CDR2 208Val Ile Trp
Tyr Gly Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val Lys1 5 10
15Gly20915PRTArtificial SequenceSynthetic 19H8 VH CDR3 209Gly Gly
Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met Asp Val1 5 10
1521011PRTArtificial SequenceSynthetic 19H8 VL1 CDR1 210Arg Ala Ser
Gln Gly Ile Ser Ser Ala Leu Ala1 5 102117PRTArtificial
SequenceSynthetic 19H8 VL1 CDR2 211Asp Ala Ser Ser Leu Glu Ser1
52129PRTArtificial SequenceSynthetic 19H8 VL1 CDR3 212Gln Gln Phe
Asn Ser Tyr Pro Gln Thr1 521311PRTArtificial SequenceSynthetic 19H8
VL2 CDR1 213Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala1 5
102147PRTArtificial SequenceSynthetic 19H8 VL2 CDR2 214Asp Ala Ser
Asn Arg Ala Thr1 52159PRTArtificial SequenceSynthetic 19H8 VL2 CDR3
215Gln Gln Arg Ser Asn Trp Pro Leu Thr1 5216449PRTArtificial
SequenceSynthetic 19H8 (VH + G2(C219S)) or 19H8-IgG2- C219S 216Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Met 35 40 45Ala Val Ile Trp Tyr Gly Gly Ser Asn Lys Phe Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Ser Leu Ser65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ala Met Val Arg Gly Val
Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val
Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170
175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr
Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
Thr Val Glu Arg 210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro
Ala Pro Pro Val Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg 290 295
300Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly217450PRTArtificial SequenceSynthetic 19H8
(VH + G2.g1) or 19H8-IgG2-IgG1 217Gln Val Gln Leu Val Glu Ser Gly
Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val Ile Trp Tyr
Gly Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Asp Asn Ser Lys Asn Ser Leu Ser65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230
235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450218449PRTArtificial SequenceSynthetic 19H8 (VH + G2.g1.1) or
19H8-IgG2- IgG1.1 218Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val
Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val Ile Trp Tyr Gly Gly Ser
Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Ser Leu Ser65 70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys Lys
Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr Ile
Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350Thr
Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 355 360
365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro
Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val
Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn His Tyr
Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly219450PRTArtificial SequenceSynthetic 19H8 (VH +
G2(C219S).g1) or 19H8- IgG2-C219S-IgG1 219Gln Val Gln Leu Val Glu
Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His Trp
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val Ile
Trp Tyr Gly Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Ser Leu Ser65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met
Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala
Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val
Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr
Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200
205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg
210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu
Leu Gly225 230 235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310 315
320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440
445Pro Gly 450220449PRTArtificial SequenceSynthetic 19H8 (VH +
G2(C219S).g1.1) or 19H8- IgG2-C219S-IgG1.1 220Gln Val Gln Leu Val
Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 20 25 30Gly Met His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Ala Val
Ile Trp Tyr Gly Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Ser Leu Ser65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Ala Met Val Arg Gly Val Tyr Tyr Tyr Gly Met
Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala
Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val
Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr
Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200
205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg
210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val
Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys305 310 315
320Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu
325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn
Gln Val Ser Leu 355 360 365Thr Cys Leu Val
Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser
Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val385 390 395
400Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
405 410 415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val
Met His 420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu
Ser Leu Ser Pro 435 440 445Gly221453PRTArtificial SequenceSynthetic
6G10.IgG1 (VH + IgG1) 221Gln Val Gln Leu Val Glu Ser Gly Gly Asp
Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser
Gly Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala
Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Thr Trp Tyr Ala Gly
Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly
Gly Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp 100 105 110Val
Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly
130 135 140Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser
Leu Gly Thr Gln Thr Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro
Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys
Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu225 230 235
240Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp
245 250 255Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val
Val Asp 260 265 270Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp
Tyr Val Asp Gly 275 280 285Val Glu Val His Asn Ala Lys Thr Lys Pro
Arg Glu Glu Gln Tyr Asn 290 295 300Ser Thr Tyr Arg Val Val Ser Val
Leu Thr Val Leu His Gln Asp Trp305 310 315 320Leu Asn Gly Lys Glu
Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 325 330 335Ala Pro Ile
Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro
Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360
365Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile
370 375 380Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr
Lys Thr385 390 395 400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe
Phe Leu Tyr Ser Lys 405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln
Gln Gly Asn Val Phe Ser Cys 420 425 430Ser Val Met His Glu Ala Leu
His Asn His Tyr Thr Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450222453PRTArtificial SequenceSynthetic 6G10.IgG1.1 (VH + IgG1.1)
222Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg1
5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr
Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ala Val Thr Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly
Leu Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr
Val Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe
Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly 130 135 140Gly Thr Ala
Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155
160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr
165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser
Ser Val 180 185 190Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr
Tyr Ile Cys Asn 195 200 205Val Asn His Lys Pro Ser Asn Thr Lys Val
Asp Lys Arg Val Glu Pro 210 215 220Lys Ser Cys Asp Lys Thr His Thr
Cys Pro Pro Cys Pro Ala Pro Glu225 230 235 240Ala Glu Gly Ala Pro
Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 245 250 255Thr Leu Met
Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 260 265 270Val
Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 275 280
285Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn
290 295 300Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln
Asp Trp305 310 315 320Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser
Asn Lys Ala Leu Pro 325 330 335Ser Ser Ile Glu Lys Thr Ile Ser Lys
Ala Lys Gly Gln Pro Arg Glu 340 345 350Pro Gln Val Tyr Thr Leu Pro
Pro Ser Arg Glu Glu Met Thr Lys Asn 355 360 365Gln Val Ser Leu Thr
Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 370 375 380Ala Val Glu
Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr385 390 395
400Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys
405 410 415Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe
Ser Cys 420 425 430Ser Val Met His Glu Ala Leu His Asn His Tyr Thr
Gln Lys Ser Leu 435 440 445Ser Leu Ser Pro Gly
450223214PRTArtificial SequenceSynthetic 6G10.IgG1 (VL + CL) 223Ala
Ile Gln Leu Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Ala
20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe
Asn Ser Tyr Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys Arg Thr Val Ala Ala 100 105 110Pro Ser Val Phe Ile Phe Pro Pro
Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125Thr Ala Ser Val Val Cys
Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140Lys Val Gln Trp
Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln145 150 155 160Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170
175Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr
180 185 190Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr
Lys Ser 195 200 205Phe Asn Arg Gly Glu Cys 210224449PRTArtificial
SequenceSynthetic 6G10 (VH + G2(C219S)) or 6G10-IgG2- C219S 224Gln
Val Gln Leu Val Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr
20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ala Val Thr Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly Leu
Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val
Thr Val Ser Ser Ala Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro
Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala
Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val
Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170
175Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val
180 185 190Val Thr Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr
Cys Asn 195 200 205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys
Thr Val Glu Arg 210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro
Ala Pro Pro Val Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro
Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu
Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu
Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn
Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg 290 295
300Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn Gly
Lys305 310 315 320Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro
Ala Pro Ile Glu 325 330 335Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro
Arg Glu Pro Gln Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu
Met Thr Lys Asn Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly
Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly
Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Met385 390 395 400Leu
Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410
415Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
420 425 430Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu
Ser Pro 435 440 445Gly225450PRTArtificial SequenceSynthetic 6G10
(VH + G2.g1) or 6G10-IgG2-IgG1 225Gln Val Gln Leu Val Glu Ser Gly
Gly Asp Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala
Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg
Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Thr Trp Tyr
Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala
Arg Gly Gly Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys
115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr
Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr
Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala
Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser
Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser
Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His
Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys
Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly225 230
235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu
Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp
Val Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val
Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr Lys Pro Arg Glu
Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val Ser Val Leu Thr
Val Leu His Gln Asp Trp Leu Asn Gly305 310 315 320Lys Glu Tyr Lys
Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 325 330 335Glu Lys
Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 340 345
350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser
355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala
Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp Gly Ser Phe Phe
Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser Arg Trp Gln Gln
Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His Glu Ala Leu His
Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440 445Pro Gly
450226449PRTArtificial SequenceSynthetic 6G10 (VH + G2.g1.1) or
6G10-IgG2- IgG1.1 226Gln Val Gln Leu Val Glu Ser Gly Gly Asp Val
Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly
Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro
Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Thr Trp Tyr Ala Gly Ser
Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser
Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser
Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Gly
Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met Asp 100 105 110Val Trp
Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys 115 120
125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu
130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro
Glu Pro145 150 155 160Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr
Ser Gly Val His Thr 165 170 175Phe Pro Ala Val Leu Gln Ser Ser Gly
Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr Val Pro Ser Ser Asn
Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200 205Val Asp His Lys Pro
Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg 210 215 220Lys Cys Cys
Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val Ala Gly225 230 235
240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser
His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly
Val Glu Val His 275 280 285Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln
Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser Val Leu Thr Val Leu
His Gln Asp Trp Leu Asn Gly Lys305 310 315 320Glu Tyr Lys Cys
Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu 325 330 335Lys Thr
Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345
350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val
Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr
Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp Gly Ser Phe Phe Leu
Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg Trp Gln Gln Gly
Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu Ala Leu His Asn
His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly227450PRTArtificial SequenceSynthetic 6G10 (VH +
G2(C219S).g1) or 6G10- IgG2-C219S-IgG1 227Gln Val Gln Leu Val Glu
Ser Gly Gly Asp Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser
Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His Trp
Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val Thr
Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys Gly
Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met
Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala
Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val
Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr
Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200
205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg
210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Glu Leu
Leu Gly225 230 235 240Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro
Lys Asp Thr Leu Met 245 250 255Ile Ser Arg Thr Pro Glu Val Thr Cys
Val Val Val Asp Val Ser His 260 265 270Glu Asp Pro Glu Val Lys Phe
Asn Trp Tyr Val Asp Gly Val Glu Val 275 280 285His Asn Ala Lys Thr
Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 290 295 300Arg Val Val
Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly305 310 315
320Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile
325 330 335Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro
Gln Val 340 345 350Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys
Asn Gln Val Ser 355 360 365Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro
Ser Asp Ile Ala Val Glu 370 375 380Trp Glu Ser Asn Gly Gln Pro Glu
Asn Asn Tyr Lys Thr Thr Pro Pro385 390 395 400Val Leu Asp Ser Asp
Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val 405 410 415Asp Lys Ser
Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 420 425 430His
Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 435 440
445Pro Gly 450228449PRTArtificial SequenceSynthetic 6G10 (VH +
G2(C219S).g1.1) or 6G10- IgG2-C219S-IgG1.1 228Gln Val Gln Leu Val
Glu Ser Gly Gly Asp Val Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu
Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Thr Tyr 20 25 30Gly Met His
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ala Val
Thr Trp Tyr Ala Gly Ser Asn Lys Phe Tyr Ala Asp Ser Val 50 55 60Lys
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr65 70 75
80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
85 90 95Ala Arg Gly Gly Ser Met Val Arg Gly Leu Tyr Tyr Tyr Gly Met
Asp 100 105 110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala
Ser Thr Lys 115 120 125Gly Pro Ser Val Phe Pro Leu Ala Pro Cys Ser
Arg Ser Thr Ser Glu 130 135 140Ser Thr Ala Ala Leu Gly Cys Leu Val
Lys Asp Tyr Phe Pro Glu Pro145 150 155 160Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser Gly Val His Thr 165 170 175Phe Pro Ala Val
Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val 180 185 190Val Thr
Val Pro Ser Ser Asn Phe Gly Thr Gln Thr Tyr Thr Cys Asn 195 200
205Val Asp His Lys Pro Ser Asn Thr Lys Val Asp Lys Thr Val Glu Arg
210 215 220Lys Ser Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val
Ala Gly225 230 235 240Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
Asp Thr Leu Met Ile 245 250 255Ser Arg Thr Pro Glu Val Thr Cys Val
Val Val Asp Val Ser His Glu 260 265 270Asp Pro Glu Val Lys Phe Asn
Trp Tyr Val Asp Gly Val Glu Val His 275 280 285Asn Ala Lys Thr Lys
Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300Val Val Ser
Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys305 310 315
320Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ser Ser Ile Glu
325 330 335Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln
Val Tyr 340 345 350Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn
Gln Val Ser Leu 355 360 365Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser
Asp Ile Ala Val Glu Trp 370 375 380Glu Ser Asn Gly Gln Pro Glu Asn
Asn Tyr Lys Thr Thr Pro Pro Val385 390 395 400Leu Asp Ser Asp Gly
Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415Lys Ser Arg
Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430Glu
Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440
445Gly
* * * * *
References